BACKGROUND At the end of December 2019,the world faced severe acute respiratory syndrome-coronavirus 2(SARS-CoV-2),which led to the outbreak of coronavirus disease 2019(COVID-19),associated with respiratory issues.Thi...BACKGROUND At the end of December 2019,the world faced severe acute respiratory syndrome-coronavirus 2(SARS-CoV-2),which led to the outbreak of coronavirus disease 2019(COVID-19),associated with respiratory issues.This virus has shown significant challenges,especially for senior citizens,patients with other underlying illnesses,or those with a sedentary lifestyle.Serological tests conducted early on have helped identify how the virus is transmitted and how to curb its spread.The study hypothesis was that the rapid serological test for SARS-CoV-2 antibodies could indicate the immunoreactive profile during the COVID-19 pandemic in a university population.AIM To conduct active surveillance for serological expression of anti-SARS-CoV-2 antibodies in individuals within a university setting during the COVID-19 pandemic.METHODS This sectional study by convenience sampling was conducted in a large university in Niteroi-RJ,Brazil,from March 2021 to July 2021.The study population consisted of students,faculty,and administrative staff employed by the university.A total of 3433 faculty members,60703 students,and 3812 administrative staff were invited to participate.Data were gathered through rapid serological tests to detect immunoglobulin(Ig)M and IgG against SARS-CoV-2.Theχ²or Fisher's exact test was used to conduct statistical analysis.A 0.20 significance level was adopted for variable selection in a multiple logistic regression model to evaluate associations.RESULTS A total of 1648 individuals were enrolled in the study.The proportion of COVID-19 positivity was 164/1648(9.8%).The adjusted logistic model indicate a positive association between the expression of IgM or IgG and age[odds ratio(OR)=1.16,95%CI:1.02-1.31](P<0.0024),individuals who had been in contact with a COVID-19-positive case(OR=3.49,95%CI:2.34-5.37)(P<0.001),those who had received the COVID-19 vaccine(OR=2.33,95%CI:1.61-3.35)(P<0.001)and social isolation(OR=0.59,95%CI:0.41-0.84)(P<0.004).The likelihood of showing a positive result increased by 16%with every ten-year increment.Conversely,adherence to social distancing measures decreased the likelihood by 41%.CONCLUSION These findings evidenced that the population became more exposed to the virus as individuals discontinued social distancing practices,thereby increasing the risk of infection for themselves.展开更多
Malaria continues to pose a significant global health challenge despite a significant achievement in control and elimination in certain areas.Accurate and timely diagnosis is crucial for effective disease management a...Malaria continues to pose a significant global health challenge despite a significant achievement in control and elimination in certain areas.Accurate and timely diagnosis is crucial for effective disease management and control,and finally leading to elimination.However,microscopy and rapid diagnostic tests(RDTs)have traditionally been the primary malaria diagnostic tools used globally,with certain shortcomings,including their limited sensitivity,specificity,and inability to identify asymptomatic infections.Serological markers have emerged as promising alternatives in malaria serosurveillance,particularly in countries where targets have already been set for elimination.This review highlights the advantages of serological markers over conventional diagnostic techniques and discusses some of the most promising serological markers against Plasmodium species-specific antigens.The implementation of serosurveillance,coupled with the utilization of these serological markers represents a transformative shift in malaria surveillance.By capitalizing on the immune memory of individuals,serosurveillance also enables the identification of recent and past infections.This approach is particularly valuable in low-transmission settings and for tracking changes in malaria prevalence over time.While recognizing the use of serological markers across various global contexts,this review predominantly emphasizes their significance within the framework of India.展开更多
In order to survey the infectious situation of canine coronavirus (CCV) in giant panda population, a virus neutralization test detecting specific antibodies against CCV in giant panda抯 sera was established by using t...In order to survey the infectious situation of canine coronavirus (CCV) in giant panda population, a virus neutralization test detecting specific antibodies against CCV in giant panda抯 sera was established by using two-fold dilutions of serum and 100 TCID50 of the virus. The 62 sera samples of giant pandas, which were gathered from zoos and reserve region of Sichuan Province, China were detected. The neutralization antibody titer of 1:4 was recognized as the positive criterion, 8 sera samples were detected to be positive, and the positive rate was 12.9%. The titers of neutralizing antibody ranged from 1:8 to 1:32. It was the first comprehensive investigation on neutralization antibodies against CCV in giant panda population in China. The results of study showed that the infection of CCV in giant panda population was universal, which has posed a threat to the health of giant panda. Therefore, it is incumbent on us to study safe and effective vaccines to protect giant panda against CCV infection.展开更多
Porcine deltacoronavirus(PDCoV)is a swine enteropathogenic CoV that causes severe vomiting,diarrhea and dehydration in suckling piglets,leading to economic losses in the swine industry.There is a great need for a conv...Porcine deltacoronavirus(PDCoV)is a swine enteropathogenic CoV that causes severe vomiting,diarrhea and dehydration in suckling piglets,leading to economic losses in the swine industry.There is a great need for a convenient method to detect circulating antibodies and help in accurate diagnosis and disease control.Previously,we demonstrated that a unique PDCoV accessory protein,NS6,is expressed during PDCoV infection in pigs and is incorporated into PDCoV virions;thus,we deduced that NS6 is likely an immunogenic target that can be used for the diagnosis of PDCoV infection.In this study,we first confirmed that NS6 is immunogenic in PDCoV-infected pigs by perform-ing a serum western blot.Furthermore,we developed a novel NS6-based indirect enzyme-linked immunosorbent assay(iELISA)method and compared it to an established S1-based iELISA for the survey of anti-PDCoV IgG or IgA in pigs of different ages in China.The NS6-iELISA has high specificity for the detection of IgG antibodies and no cross-reactivity with other porcine enteric CoVs(transmissible gastroenteritis coronavirus,porcine epidemic diarrhea virus,or swine acute diarrhea syndrome coronavirus).This NS6 serology-based method has great sensitivity and good repeatability,making it a new and cost-saving option for the rapid diagnosis and immunosurveillance of PDCoV,which may also be important for the prevention and control of deltacoronavirus-related infection in pigs and other animals.展开更多
[ Objective] To investigate the prevalence of Mycoplasma oviovipneurnoniae in Qinghai Province. [ Method] With positive indirect hemagglutination test kit for detecting antibodies against Mycoplasma oviovipneumoniae, ...[ Objective] To investigate the prevalence of Mycoplasma oviovipneurnoniae in Qinghai Province. [ Method] With positive indirect hemagglutination test kit for detecting antibodies against Mycoplasma oviovipneumoniae, 965 sheep sera and 208 goat sera were collected in Qinghai Province from 2006 to 2008 and detected. [ Result ] The positive rate of sheep sera and goat sera was 30.4% and 19.7%, respectively. The posi- tive rate of sheep sera collected from different regions ranged from 19.7% to 40.2%, and that of goat sere ranged from 6.6% to 22.2%. In addition, the incidence in winter and spring was higher than that in summer and autumn. [ Conclusion ] The infection rate of Mycoplasrna ovipneumoniae should be higher in Qinghai region than in other western regions, so it is necessary to strengthen the prevention and control of this disease.展开更多
Correction to“Freire de Melo F,Martins Oliveira Diniz L,Nélio Januário J,Fernando Gonçalves Ferreira J,Dórea RSDM,de Brito BB,Marques HS,Lemos FFB,Silva Luz M,Rocha Pinheiro SL,de Magalhães Q...Correction to“Freire de Melo F,Martins Oliveira Diniz L,Nélio Januário J,Fernando Gonçalves Ferreira J,Dórea RSDM,de Brito BB,Marques HS,Lemos FFB,Silva Luz M,Rocha Pinheiro SL,de Magalhães Queiroz DM.Performance of a serological IgM and IgG qualitative test for COVID-19 diagnosis:An experimental study in Brazil.World J Exp Med 2022;12(5):100-103[PMID:36196438 DOI:10.5493/wjem.v12.i5.100]”.In this article,we identified an issue with the“Acknowledgments”section.Here,we then provide a recognition section for our supporting institutions.展开更多
The panel of serologic markers for inflammatory bowel diseases （IBD） is rapidly expanding. Although antiSaccharornyces cerev/siae antibodies （ASCA） and atypical perinuclear antineutrophil cytoplasmic antibodies （...The panel of serologic markers for inflammatory bowel diseases （IBD） is rapidly expanding. Although antiSaccharornyces cerev/siae antibodies （ASCA） and atypical perinuclear antineutrophil cytoplasmic antibodies （P-ANCA） remain the most widely investigated, an increasing amount of experimental data is available on newly discovered antibodies directed against various microbial antigens. The role of the assessment of various antibodies in the current IBD diagnostic algorithm is often questionable due to their limited sensitivity. In contrast, the association of serologic markers with disease behavior and phenotype is becoming increasingly well-established. An increasing number of observations confirms that patients with Crohn＇s disease expressing multiple serologic markers at high titers are more likely to have complicated small bowel disease （e.g. stricture and/or perforation） and are at higher risk for surgery than those without, or with low titers of antibodies. Creating homogenous disease sub-groups based on serologic response may help develop more standardized therapeutic approaches and may help in a better understanding of the pathomechanism of IBD. Further prospective clinical studies are needed to establish the clinical role of serologic tests in IBD.展开更多
Objective To investigate the relationship between maternal peripheral blood mononuclear cells (PBMC) hepatitis B virus(HBV) covalenty closed circular deoxyribonucleic acid(cccDNA) and other HBV serological markers and...Objective To investigate the relationship between maternal peripheral blood mononuclear cells (PBMC) hepatitis B virus(HBV) covalenty closed circular deoxyribonucleic acid(cccDNA) and other HBV serological markers and its effects on HBV intrauterine transmission. Methods We enrolled 290 newborns and their hepatitis B surface antigen(HBsAg) positive mothers. HBV cccDNA in PBMC and HBV DNA in serum were detected by a real‐time PCR‐TaqM an probe while HBV serological markers were detected with an electrochemiluminescence immunoassay. Results There was a positive correlation between the levels of PBMC HBV cccD NA and serum HBV DNA and HBeA g(r = 0.436 and 0.403, P < 0.001). The detection rate of pattern A [‘HBsA g(+), HBeA g(+), and anti‐HBc(+)’] was significantly higher in the PBMC HBV cccD NA positive group than in the control group(χ^2 = 48.48, P < 0.001). There was a significant association between HBV intrauterine transmission and PBMC HBV cccD NA(χ^2 = 9.28, P = 0.002). In the presence of serum HBV DNA, HBeA g, and PBMC HBV cccD NA, the risk of HBV intrauterine transmission was three times higher(OR = 3.69, 95% CI: 1.30‐10.42) than that observed in their absence. The risk of HBV intrauterine transmission was the greatest(OR = 5.89, 95% CI: 2.35‐14.72) when both PBMC HBV cccD NA and pattern A were present. A Bayesian network model showed that maternal PBMC HBV cccD NA was directly related to HBV intrauterine transmission. Conclusion PBMC HBV cccDNA may be a direct risk factor for HBV intrauterine transmission. Our study suggests that serological markers could be combined with PBMC‐related markers in prenatal testing.展开更多
Citrus yellow vein clearing virus （CYVCV） is considered as the causal agent of Citrus yellow vein clearing disease and belongs to the genus Mandarivirus in the family Alphaflexiviridae. Capsid protein （CP） of CYVC...Citrus yellow vein clearing virus （CYVCV） is considered as the causal agent of Citrus yellow vein clearing disease and belongs to the genus Mandarivirus in the family Alphaflexiviridae. Capsid protein （CP） of CYVCV Chongqing isolate （CYVCV- CQ） was produced using a prokaryotic expression system and used as the immunogen for monoclonal antibody （MAb） production. Four highly specific and sensitive murine MAbs and one polyclonal antibody were prepared in this study. Titers of the four MAbs in ascites fluids ranged from 10-6 to 10-7 as determined by indirect enzyme-linked immunosorbent assay （ELISA）. Three serological assays, including dot enzyme-linked immunosorbent assay （dot-ELISA）, tissue blot-ELISA, and double-antibody sandwich （DAS）-ELISA, were developed for quick and reliable detections of CYVCV in citrus samples. The developed dot-ELISA and DAS-ELISA methods could detect CYVCV in the infected citrus leaf crude extracts diluted at 1：2 560 and 1：10 240 （w/v, g mL^-1）, respectively. The detection result of 125 citrus leaf samples collected from citrus groves in Yunnan Province and Chongqing Municipality of China showed that approximately 36% samples were positive for CYVCV. This virus was, however, not＇detected in any sample collected from Zhejiang or Jiangxi Province, China.展开更多
The immense patient number caused by coronavirus disease 2019(COVID-19)global pandemic brings the urge for more knowledge about its immunological features,including the profile of basic immune parameters.In this study...The immense patient number caused by coronavirus disease 2019(COVID-19)global pandemic brings the urge for more knowledge about its immunological features,including the profile of basic immune parameters.In this study,eighty-eight reported COVID-19 patients in Wuhan were recruited from January to February,2020,including 32 severe/critical cases and56 mild/moderate cases.Their mean age was 56.43 years(range 17–83)and gender ratio(male/female)was 43:45.We tested SARS-CoV-2 RNA with commercial kits,investigated the level of serologic IgM and IgG antibodies against severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)using magnetic particle chemiluminescence immunoassays,and compared the results of serologic tests and nucleic acid test(NAT).Among 88 patients,95.45%were confirmed as positive by the combination of NAT and antibody test,which was significantly higher(P<0.001)than by single nucleic acid test(73.86%)or serologic test(65.91%).Then the correlation between temporal profile and the level of antibody response was analyzed.It showed that seroconversion started on day 5 after disease onset and IgG level was rose earlier than IgM.Comparison between patients with different disease severity suggested early seroconversion and high antibody titer were linked with less severe clinical symptoms.These results supported the combination of serologic testing and NAT in routine COVID-19 diagnosis and provided evidence on the temporal profile of antibody response in patients with different disease severity.展开更多
Potato virus M(PVM) is one of the common and economically important potato viruses in potato-growing regions worldwide. To investigate and control this viral disease, efficient and specific detection techniques are ne...Potato virus M(PVM) is one of the common and economically important potato viruses in potato-growing regions worldwide. To investigate and control this viral disease, efficient and specific detection techniques are needed. In this study, PVM virions were purified from infected potato plants and used as the immunogen to produce hybridomas secreting PVM-specific monoclonal antibodies(MAbs). Four highly specific and sensitive murine MAbs, i.e., 1 E1, 2 A5, 8 A1 and 17 G8 were prepared through a conventional hybridoma technology. Using these four MAbs, we have developed an antigen-coated plate(ACP)-ELISA, a dot-ELISA and a Tissue print-ELISA for detecting PVM infection in potato plants and tubers. PVM could be detected in infected potato plant tissue crude extracts diluted at 1:10 240(w/v, g mL^(–1)) by the dot-ELISA or at 1:163 840(w/v, g mL^(–1)) by the ACP-ELISA. The Tissue print-ELISA is the quickest and easiest assay among the three established serological assays and is more suitable for onsite large-scale sample detection. Detection results of the field-collected samples showed that PVM is currently widespread in the Yunnan and the Heilongjiang provinces in China. The field sample test results of the developed serological assays were supported by the results from RT-PCR and DNA sequencing. We consider that the newly established ACP-ELISA, dot-ELISA and Tissue print-ELISA can benefit PVM detection in potato plant and tuber samples and field epidemiological studies of PVM. These assays can also facilitate the production of virus-free seed potatoes and breeding for PVM-resistant potato cultivars, leading to the successful prevention of this potato viral disease.展开更多
Rice stripe mosaic virus(RSMV) is a rhabdovirus recently found in southern part of China and can cause severe reduction in rice production. To establish serological methods for RSMV epidemiological studies and to esta...Rice stripe mosaic virus(RSMV) is a rhabdovirus recently found in southern part of China and can cause severe reduction in rice production. To establish serological methods for RSMV epidemiological studies and to establish a control strategy for this virus, we first purified RSMV virions from infected rice plants and then used them as an immunogen to produce four RSMV-specific monoclonal antibodies(MAbs)(i.e.,1D4, 4A8, 8E4 and 11F11). With these MAbs, we have developed a highly specific and sensitive antigen-coated plate enzyme-linked immunosorbent assay(ACP-ELISA), a Dot-ELISA and a Tissue print-ELISA for rapid detections of RSMV infection in rice plants or in leafhoppers. Our results showed that RSMV can be readily detected in RSMV-infected rice plant tissue crude extracts diluted at 1:20,971,520(w/v, g/m L)through ACP-ELISA or diluted at 1:327,680(w/v, g/m L) through Dot-ELISA. Both ACP-ELISA and Dot-ELISA can also be used to detect RSMV infection in individual RSMV viruliferous leafhopper(Recilia dorsalis) homogenate diluted at 1:307,200 and 1:163,840(individual leafhopper/l L), respectively. Detection of RSMV infection in field-collected rice samples or in RSMV viruliferous leafhoppers indicated that the three serological methods can produce same results with that produced by RT-PCR(19 of the 33 rice samples and 5 of the 16 leafhoppers were RSMV-positive). We consider that the four MAbs produced in this study are very specific and sensitive, and the three new serological methods are very useful for detections of RSMV infection in rice plants or in leafhoppers and the establishment of the disease control strategies.展开更多
BACKGROUND Currently, it is difficult to predict the complications of children at the early stage of sepsis. Brighton pediatric early warning score(PEWS) is a disease risk assessment system that is simple and easy to ...BACKGROUND Currently, it is difficult to predict the complications of children at the early stage of sepsis. Brighton pediatric early warning score(PEWS) is a disease risk assessment system that is simple and easy to operate, which has good sensitivity and specificity in disease recognition among children. Because detection indicators vary widely in children, a single indicator is difficult to assess the posttreatment status of children with sepsis.AIM To investigate the relationship between serological markers, Brighton PEWS, and death in children with sepsis after treatment.METHODS A total of 205 children diagnosed with sepsis at our hospital were enrolled. The baseline data, serum scores, and PEWS scores were recorded. In the nested casecontrol study, children who died during the study period were included in an observation group. According to the matching principle, the children who were not dead in the same cohort were included in a control group. The influencing factors of death in children with sepsis after treatment and the value of each evaluation index in predicting the prognosis of children were analyzed.RESULTS A total of 96 children were enrolled in the study, including 48 each in the observation group and the control group. Multivariate logistic regression analysis indicated that antibacterial treatments within 1 h(P = 0.017), shock(P = 0.044),multiple organ dysfunction syndrome(MODS)(P = 0.027), serum procalcitonin(PCT)(P = 0.047), serum albumin(ALB)(P = 0.024), and PEWS(P = 0.012) were independent risk factors for the death of children with sepsis. The area under the curve of the combination of ALB, PCT, and PEWS to predict the death in children with sepsis was the highest(0.908).CONCLUSION Antibacterial treatments within 1 h, shock, MODS, PCT, ALB, and PEWS are independent risk factors for the death of children with sepsis. The predictive accuracy of the combination of PCT, ALB, and PEWS for the prognosis of children with sepsis is the best.展开更多
Serological biomarkers in inflammatory bowel disease (IBD) are a rapidly expanding list of non-invasive tests for objective assessments of disease activity, early diagnosis, prognosis evaluation and surveillance. This...Serological biomarkers in inflammatory bowel disease (IBD) are a rapidly expanding list of non-invasive tests for objective assessments of disease activity, early diagnosis, prognosis evaluation and surveillance. This review summarizes both old and new biomarkers in IBD, but focuses on the development and character-ization of new serological biomarkers (identifi ed since 2007). These include fi ve new anti-glycan antibodies, anti-chitobioside IgA (ACCA), anti-laminaribioside IgG (ALCA), anti-manobioside IgG (AMCA), and antibod-ies against chemically synthesized (∑) two major oligomannose epitopes, Man α-1,3 Man α-1,2 Man (∑Man3) and Man α-1,3 Man α-1,2 Man α-1,2 Man (∑Man4). These new biomarkers serve as valuable complementary tools to existing biomarkers not only in differentiating Crohn's disease (CD), ulcerative colitis (UC), normal and other non-IBD gut diseases, but also in predicting disease involvement (ileum vs colon), IBD risk (as subclinical biomarkers), and disease course (risk of complication and surgery). Interestingly, the prevalence of the antiglycan antibodies, including anti-Saccharomyces cerevisiae antibodies (ASCA), ALCA and AMCA, was found to be associated with single nucleotide polymorphisms (SNPs) of IBD susceptible genes such as NOD2/CARD15, NOD1/CARD4, toll-likereceptors (TLR) 2 and 4, and β-defensin-1. Further-more, a gene dosage effect was observed: anti-glycan positivity became more frequent as the number of NOD2/CARD15 SNPS increased. Other new serum/ plasma IBD biomarkers reviewed include ubiquitination factor E4A (UBE4A), CXCL16 (a chemokine), resistin, and apolipoprotein A-IV. This review also discusses the most recent studies in IBD biomarker discovery by the application of new technologies such as proteomics, fourier transform near-infrared spectroscopy, and mul-tiplex enzyme-linked immunosorbent assay (ELISA)'s (with an emphasis on cytokine/chemokine profiling). Finally, the prospects of developing more clinically use-ful novel diagnostic algorithms by incorporating new technologies in serological biomarker profiling and integrating multiple biomarkers with bioinformatics analysis/modeling are also discussed.展开更多
Pepino mosaic virus(PepMV)causes severe disease in tomato and other Solanaceous crops around globe.To effectively study and manage this viral disease,researchers need new,sensitive,and high-throughput approaches for v...Pepino mosaic virus(PepMV)causes severe disease in tomato and other Solanaceous crops around globe.To effectively study and manage this viral disease,researchers need new,sensitive,and high-throughput approaches for viral detection.In this study,we purified PepMV particles from the infected Nicotiana benthamiana plants and used virions to immunize BALB/c mice to prepare hybridomas secreting anti-PepMV monoclonal antibodies(mAbs).A panel of highly specific and sensitive murine mAbs(15B2,8H6,23D11,20D9,3A6,and 8E3)could be produced through cell fusion,antibody selection,and cell cloning.Using the mAbs as the detection antibodies,we established double antibody sandwich enzyme-linked immunosorbent assay(DAS-ELISA),Dot-ELISA,and Tissue print-ELISA for detecting PepMV infection in tomato plants.Resulting data on sensitivity analysis assays showed that both DAS-ELISA and Dot-ELISA can efficiently monitor the virus in PepMV-infected tissue crude extracts when diluted at 1:1310720 and 1:20480(weight/volume ratio(w/v),g/mL),respectively.Among the three methods developed,the Tissue print-ELISA was found to be the most practical detection technique.Survey results from field samples by the established serological approaches were verified by reverse transcription polymerase chain reaction(RT-PCR)and DNA sequencing,dem on strati ng all three serological methods are reliable and effective for monitoring PepMV.An ti-PepMV mAbs and the newly developed DAS-ELISA,Dot-ELISA,and Tissue print-ELISA can benefit PepMV detection and field epidemiological study,and management of this viral disease,which is already widespread in tomato plants in Yunnan Province of China.展开更多
Objective:To evaluate the Anaplasma phagocytophilum(A.phagocytophilum),Ehrlichia canis(E.canis,Dirofilaria immitis(D.immitis)(canine heartworm),Borrelia burgdorferi(B.burgdorferi)infections in countryside dogs from Yu...Objective:To evaluate the Anaplasma phagocytophilum(A.phagocytophilum),Ehrlichia canis(E.canis,Dirofilaria immitis(D.immitis)(canine heartworm),Borrelia burgdorferi(B.burgdorferi)infections in countryside dogs from Yunnan,Hainan and Anhui provinces.Methods:Serum samples were collected from 26 dogs in Yunnan.Hainan and Anhui provinces.The samples were tested using a commercial ELISA rapid diagnostic assay kit(SNAP^(?)4Dx^(?);IDEXX Laboratories,Inc.U.S.A.).Meaiiwliile,indirect immunofluorescence assay(IFA)recommended by WHO was conducted to delect IgG to A.phagocytophilum.Two methods were analyzed and compared.Results:The number of serologically positive dogs for IgG to A.phagocytophilum was only 2which was from Hainan province and none of the 26 dogs responded positive for E.canu.D.immitis(canine heartworm,and B.burgdorferi by ELISA rapid diagnostic method.The number of serologically positive dogs for IgG to A.phagocytophilum was 13(50%)by IFA method.Data of the two methods were analyzed by statistical software and the difference was statistically significant(P=0.002).Conclusions:It can be concluded that IFA method was more sensitive than ELISA rapid diagnostic method.However,we need conduct further and intensive epidemiology survey on tick-born diseases pathogens including.4.phagocytophilum,E.canis,D.immitis(canine heartworm),and B.burgdorferi which have public health significance.展开更多
Many flaviviruses are emerging and reemerging pathogens, such as West Nile virus （WNV）, dengue virus （DENV）, yellow fever virus （YFV）, and Japanese encephalitis virus. Serological assay is the dominant method fo...Many flaviviruses are emerging and reemerging pathogens, such as West Nile virus （WNV）, dengue virus （DENV）, yellow fever virus （YFV）, and Japanese encephalitis virus. Serological assay is the dominant method for diagnosis of flavivirus infections in human. Because antibodies generated during flavivirus infections cross-react with other flavivirus members, plaque reduction neutralization test （PRNT） is the only available assay to determine the infecting flavivirus type. Since PRNT requires culturing raw viruses, it must be performed in biosafety levet-3 or level-4 containment for many flaviviruses, and takes more than ten days to complete. To overcome these problems, we have developed flavivirus viral-like particles （VLPs） that could be used to replace raw viruses in the neutralization assay. The VLPs were prepared by trans packaging a luciferase-reporting replicon with viral structural proteins. This novel assay involves three simple steps： （i） VLPs from a panel of flaviviruses are incubated with flavivirus-infected sera at 37℃ for 1 h; （ii）the neutralized VLPs are used to infect Vero cells; and （iii） the infected cells are measured for luciferase activities at 22 h post-infection. The virus type whose VLP is most efficiently neutralized by the serum specimen （as quantified by the luciferase activities） is the etiologic agent. As a proof-of-concept, we show that a WNV-infected mouse serum neutralized the WNV VLP more efficiently and selectively than the DENV and YFV VLPs. Our results demonstrate that the VLP neutralization assay maintains the ＂gold standard＂ of the classic PRNT; importantly, it shortens the assay time from 〉10 days to 〈1 day, and can be performed in biosafety level-2 facility.展开更多
This study aimed to investigate the serological characteristics of Ebola virus(EBOV) infection during the late phase of the Ebola outbreak in Sierra Leone. In total, 877 blood samples from 694 suspected Ebola virus di...This study aimed to investigate the serological characteristics of Ebola virus(EBOV) infection during the late phase of the Ebola outbreak in Sierra Leone. In total, 877 blood samples from 694 suspected Ebola virus disease(EVD) cases assessed from March to December 2015, were analyzed via real-time reverse transcription polymerase chain reaction(RT-PCR) for viral RNA and enzyme-linked immunosorbent assay(ELISA) and Luminex to detect antibodies against EBOV. Viral load and EBOV-specific IgM/IgG titers displayed a declining trend during March to December 2015. Viral RNA load decreased rapidly at earlier stages after disease onset, while EBOV-specific IgM and IgG still persisted in 58.1%(18/31) and 93.5%(29/31) of the confirmed EVD patients and in 3.8%(25/663) and 17.8%(118/663) of the RNA-negative suspected patients in the later phase, respectively. Dynamic analysis of longitudinally collected samples from eight EVD patients revealed typically reversed trends of declining viral load and increasing IgM and/or IgG titers in response to the EBOV infection.The present results indicate that certain populations of Sierra Leone developed immunity to an EBOV infection in the late phase of the outbreak, providing novel insights into the risk assessment of EBOV infections among human populations.展开更多
Severe fever with thrombocytopenia syndrome(SFTS),caused by SFTS virus(SFTSV)infection,was first reported in 2010 in China with an initial fatality of up to 30%.The laboratory confirmation of SFTSV infection in terms ...Severe fever with thrombocytopenia syndrome(SFTS),caused by SFTS virus(SFTSV)infection,was first reported in 2010 in China with an initial fatality of up to 30%.The laboratory confirmation of SFTSV infection in terms of detection of viral RNA or antibody levels is critical for SFTS diagnosis and therapy.In this study,a new luciferase immunoprecipitation system(LIPS)assay based on p REN2 plasmid expressing SFTSV NP gene and tagged with Renilla luciferase(Rluc),was established and used to investigate the levels of antibody responses to SFTSV.Totally 464 serum samples from febrile patients were collected in the hospital of Shaoxing City in Zhejiang Province in 2019.The results showed that 82 of the 464 patients(17.7%)had antibody response to SFTSV,which were further supported by immunofluorescence assays(IFAs).Further,q RT-PCR and microneutralization tests showed that among the 82 positive cases,15 patients had viremia,10 patients had neutralizing antibody,and one had both(totally 26 patient).However,none of these patients were diagnosed as SFTS in the hospital probably because of their mild symptoms or subclinical manifestations.All the results indicated that at least the 26 patients having viremia or neutralizing antibody were the missed diagnosis of SFTS cases.The findings suggested the occurrence of SFTS and the SFTS incidence were higher than the reported level in Shaoxing in 2019,and that LIPS may provide an alternative strategy to confirm SFTSV infection in the laboratory.展开更多
The spectrum of serological markers associated with inflammatory bowel disease (IBD) is rapidly growing. Due to frequently delayed or missed diagnoses, the application of non-invasive diagnostic tests for IBD, as well...The spectrum of serological markers associated with inflammatory bowel disease (IBD) is rapidly growing. Due to frequently delayed or missed diagnoses, the application of non-invasive diagnostic tests for IBD, as well as differentiation between ulcerative colitis (UC) and Crohn’s disease (CD), would be useful in the pediatric population. In addition, the combination of pancreatic autoantibodies and antibodies against Saccharomyces cerevisiae antibodies/perinuclear cytoplasmic antibody (pANCA) improved the sensitivity of serological markers in pediatric patients with CD and UC. Some studies suggested that age-associated differences in the patterns of antibodies may be present, particularly in the youngest children. In CD, most patients develop stricturing or perforating complications, and a significant number of patients undergo surgery during the disease course. Based on recent knowledge, serum antibodies are qualitatively and quantitatively associated with complicated CD behavior and CD-related surgery. Pediatric UC is characterized by extensive colitis and a high rate of colectomy. In patients with UC, high levels of anti-CBir1 and pANCA are associated with the development of pouchitis after ileal pouch-anal anastomosis. Thus, serologic markers for IBD can be applied to stratify IBD patients into more homogeneous subgroups with respect to disease progression. In conclusion, identification of patients at an increased risk of rapid disease progression is of great interest, as the application of early and more aggressive pharmaceutical intervention could have the potential to alter the natural history of IBD, and reduce complications and hospitalizations.展开更多
文摘BACKGROUND At the end of December 2019,the world faced severe acute respiratory syndrome-coronavirus 2(SARS-CoV-2),which led to the outbreak of coronavirus disease 2019(COVID-19),associated with respiratory issues.This virus has shown significant challenges,especially for senior citizens,patients with other underlying illnesses,or those with a sedentary lifestyle.Serological tests conducted early on have helped identify how the virus is transmitted and how to curb its spread.The study hypothesis was that the rapid serological test for SARS-CoV-2 antibodies could indicate the immunoreactive profile during the COVID-19 pandemic in a university population.AIM To conduct active surveillance for serological expression of anti-SARS-CoV-2 antibodies in individuals within a university setting during the COVID-19 pandemic.METHODS This sectional study by convenience sampling was conducted in a large university in Niteroi-RJ,Brazil,from March 2021 to July 2021.The study population consisted of students,faculty,and administrative staff employed by the university.A total of 3433 faculty members,60703 students,and 3812 administrative staff were invited to participate.Data were gathered through rapid serological tests to detect immunoglobulin(Ig)M and IgG against SARS-CoV-2.Theχ²or Fisher's exact test was used to conduct statistical analysis.A 0.20 significance level was adopted for variable selection in a multiple logistic regression model to evaluate associations.RESULTS A total of 1648 individuals were enrolled in the study.The proportion of COVID-19 positivity was 164/1648(9.8%).The adjusted logistic model indicate a positive association between the expression of IgM or IgG and age[odds ratio(OR)=1.16,95%CI:1.02-1.31](P<0.0024),individuals who had been in contact with a COVID-19-positive case(OR=3.49,95%CI:2.34-5.37)(P<0.001),those who had received the COVID-19 vaccine(OR=2.33,95%CI:1.61-3.35)(P<0.001)and social isolation(OR=0.59,95%CI:0.41-0.84)(P<0.004).The likelihood of showing a positive result increased by 16%with every ten-year increment.Conversely,adherence to social distancing measures decreased the likelihood by 41%.CONCLUSION These findings evidenced that the population became more exposed to the virus as individuals discontinued social distancing practices,thereby increasing the risk of infection for themselves.
文摘Malaria continues to pose a significant global health challenge despite a significant achievement in control and elimination in certain areas.Accurate and timely diagnosis is crucial for effective disease management and control,and finally leading to elimination.However,microscopy and rapid diagnostic tests(RDTs)have traditionally been the primary malaria diagnostic tools used globally,with certain shortcomings,including their limited sensitivity,specificity,and inability to identify asymptomatic infections.Serological markers have emerged as promising alternatives in malaria serosurveillance,particularly in countries where targets have already been set for elimination.This review highlights the advantages of serological markers over conventional diagnostic techniques and discusses some of the most promising serological markers against Plasmodium species-specific antigens.The implementation of serosurveillance,coupled with the utilization of these serological markers represents a transformative shift in malaria surveillance.By capitalizing on the immune memory of individuals,serosurveillance also enables the identification of recent and past infections.This approach is particularly valuable in low-transmission settings and for tracking changes in malaria prevalence over time.While recognizing the use of serological markers across various global contexts,this review predominantly emphasizes their significance within the framework of India.
基金This research was supported by National Science Founda-tion of China (No. 30000123) and Conversation Department of Wildlife Ani-mal & Plants of State Forestry Bureau.
文摘In order to survey the infectious situation of canine coronavirus (CCV) in giant panda population, a virus neutralization test detecting specific antibodies against CCV in giant panda抯 sera was established by using two-fold dilutions of serum and 100 TCID50 of the virus. The 62 sera samples of giant pandas, which were gathered from zoos and reserve region of Sichuan Province, China were detected. The neutralization antibody titer of 1:4 was recognized as the positive criterion, 8 sera samples were detected to be positive, and the positive rate was 12.9%. The titers of neutralizing antibody ranged from 1:8 to 1:32. It was the first comprehensive investigation on neutralization antibodies against CCV in giant panda population in China. The results of study showed that the infection of CCV in giant panda population was universal, which has posed a threat to the health of giant panda. Therefore, it is incumbent on us to study safe and effective vaccines to protect giant panda against CCV infection.
基金supported by the Zhejiang Provincial Natural Science Foundation(LZ22C180002)the Laboratory of Lingnan Modern Agriculture Project(NG2022001)the Zhejiang Provincial Key R&D Program of China(2021C02049)。
文摘Porcine deltacoronavirus(PDCoV)is a swine enteropathogenic CoV that causes severe vomiting,diarrhea and dehydration in suckling piglets,leading to economic losses in the swine industry.There is a great need for a convenient method to detect circulating antibodies and help in accurate diagnosis and disease control.Previously,we demonstrated that a unique PDCoV accessory protein,NS6,is expressed during PDCoV infection in pigs and is incorporated into PDCoV virions;thus,we deduced that NS6 is likely an immunogenic target that can be used for the diagnosis of PDCoV infection.In this study,we first confirmed that NS6 is immunogenic in PDCoV-infected pigs by perform-ing a serum western blot.Furthermore,we developed a novel NS6-based indirect enzyme-linked immunosorbent assay(iELISA)method and compared it to an established S1-based iELISA for the survey of anti-PDCoV IgG or IgA in pigs of different ages in China.The NS6-iELISA has high specificity for the detection of IgG antibodies and no cross-reactivity with other porcine enteric CoVs(transmissible gastroenteritis coronavirus,porcine epidemic diarrhea virus,or swine acute diarrhea syndrome coronavirus).This NS6 serology-based method has great sensitivity and good repeatability,making it a new and cost-saving option for the rapid diagnosis and immunosurveillance of PDCoV,which may also be important for the prevention and control of deltacoronavirus-related infection in pigs and other animals.
基金Supported by Special fund for Agricultural Biotechnology of Gansu Province ( GNSW-2005-16)Major Sci-tech Fund of Gansu Province (0702NKDA040)~~
文摘[ Objective] To investigate the prevalence of Mycoplasma oviovipneurnoniae in Qinghai Province. [ Method] With positive indirect hemagglutination test kit for detecting antibodies against Mycoplasma oviovipneumoniae, 965 sheep sera and 208 goat sera were collected in Qinghai Province from 2006 to 2008 and detected. [ Result ] The positive rate of sheep sera and goat sera was 30.4% and 19.7%, respectively. The posi- tive rate of sheep sera collected from different regions ranged from 19.7% to 40.2%, and that of goat sere ranged from 6.6% to 22.2%. In addition, the incidence in winter and spring was higher than that in summer and autumn. [ Conclusion ] The infection rate of Mycoplasrna ovipneumoniae should be higher in Qinghai region than in other western regions, so it is necessary to strengthen the prevention and control of this disease.
文摘Correction to“Freire de Melo F,Martins Oliveira Diniz L,Nélio Januário J,Fernando Gonçalves Ferreira J,Dórea RSDM,de Brito BB,Marques HS,Lemos FFB,Silva Luz M,Rocha Pinheiro SL,de Magalhães Queiroz DM.Performance of a serological IgM and IgG qualitative test for COVID-19 diagnosis:An experimental study in Brazil.World J Exp Med 2022;12(5):100-103[PMID:36196438 DOI:10.5493/wjem.v12.i5.100]”.In this article,we identified an issue with the“Acknowledgments”section.Here,we then provide a recognition section for our supporting institutions.
文摘The panel of serologic markers for inflammatory bowel diseases （IBD） is rapidly expanding. Although antiSaccharornyces cerev/siae antibodies （ASCA） and atypical perinuclear antineutrophil cytoplasmic antibodies （P-ANCA） remain the most widely investigated, an increasing amount of experimental data is available on newly discovered antibodies directed against various microbial antigens. The role of the assessment of various antibodies in the current IBD diagnostic algorithm is often questionable due to their limited sensitivity. In contrast, the association of serologic markers with disease behavior and phenotype is becoming increasingly well-established. An increasing number of observations confirms that patients with Crohn＇s disease expressing multiple serologic markers at high titers are more likely to have complicated small bowel disease （e.g. stricture and/or perforation） and are at higher risk for surgery than those without, or with low titers of antibodies. Creating homogenous disease sub-groups based on serologic response may help develop more standardized therapeutic approaches and may help in a better understanding of the pathomechanism of IBD. Further prospective clinical studies are needed to establish the clinical role of serologic tests in IBD.
基金supported by grants from the National Natural Science Foundation of China [81573212,81872677]Open Project Support by the State Key Laboratory of Infectious Disease Prevention and Control [2017SKLID306,2018SKLID310]
文摘Objective To investigate the relationship between maternal peripheral blood mononuclear cells (PBMC) hepatitis B virus(HBV) covalenty closed circular deoxyribonucleic acid(cccDNA) and other HBV serological markers and its effects on HBV intrauterine transmission. Methods We enrolled 290 newborns and their hepatitis B surface antigen(HBsAg) positive mothers. HBV cccDNA in PBMC and HBV DNA in serum were detected by a real‐time PCR‐TaqM an probe while HBV serological markers were detected with an electrochemiluminescence immunoassay. Results There was a positive correlation between the levels of PBMC HBV cccD NA and serum HBV DNA and HBeA g(r = 0.436 and 0.403, P < 0.001). The detection rate of pattern A [‘HBsA g(+), HBeA g(+), and anti‐HBc(+)’] was significantly higher in the PBMC HBV cccD NA positive group than in the control group(χ^2 = 48.48, P < 0.001). There was a significant association between HBV intrauterine transmission and PBMC HBV cccD NA(χ^2 = 9.28, P = 0.002). In the presence of serum HBV DNA, HBeA g, and PBMC HBV cccD NA, the risk of HBV intrauterine transmission was three times higher(OR = 3.69, 95% CI: 1.30‐10.42) than that observed in their absence. The risk of HBV intrauterine transmission was the greatest(OR = 5.89, 95% CI: 2.35‐14.72) when both PBMC HBV cccD NA and pattern A were present. A Bayesian network model showed that maternal PBMC HBV cccD NA was directly related to HBV intrauterine transmission. Conclusion PBMC HBV cccDNA may be a direct risk factor for HBV intrauterine transmission. Our study suggests that serological markers could be combined with PBMC‐related markers in prenatal testing.
基金supported by the Special Fund for Agro-scientific Research in the Public Interest,China(201203076-05)the National Basic Research Program of China(2014CB138400)
文摘Citrus yellow vein clearing virus （CYVCV） is considered as the causal agent of Citrus yellow vein clearing disease and belongs to the genus Mandarivirus in the family Alphaflexiviridae. Capsid protein （CP） of CYVCV Chongqing isolate （CYVCV- CQ） was produced using a prokaryotic expression system and used as the immunogen for monoclonal antibody （MAb） production. Four highly specific and sensitive murine MAbs and one polyclonal antibody were prepared in this study. Titers of the four MAbs in ascites fluids ranged from 10-6 to 10-7 as determined by indirect enzyme-linked immunosorbent assay （ELISA）. Three serological assays, including dot enzyme-linked immunosorbent assay （dot-ELISA）, tissue blot-ELISA, and double-antibody sandwich （DAS）-ELISA, were developed for quick and reliable detections of CYVCV in citrus samples. The developed dot-ELISA and DAS-ELISA methods could detect CYVCV in the infected citrus leaf crude extracts diluted at 1：2 560 and 1：10 240 （w/v, g mL^-1）, respectively. The detection result of 125 citrus leaf samples collected from citrus groves in Yunnan Province and Chongqing Municipality of China showed that approximately 36% samples were positive for CYVCV. This virus was, however, not＇detected in any sample collected from Zhejiang or Jiangxi Province, China.
基金supported by the Emergency Scientific Research Project for COVID-19 from Wuhan City(Grant No.2020020101010008)。
文摘The immense patient number caused by coronavirus disease 2019(COVID-19)global pandemic brings the urge for more knowledge about its immunological features,including the profile of basic immune parameters.In this study,eighty-eight reported COVID-19 patients in Wuhan were recruited from January to February,2020,including 32 severe/critical cases and56 mild/moderate cases.Their mean age was 56.43 years(range 17–83)and gender ratio(male/female)was 43:45.We tested SARS-CoV-2 RNA with commercial kits,investigated the level of serologic IgM and IgG antibodies against severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)using magnetic particle chemiluminescence immunoassays,and compared the results of serologic tests and nucleic acid test(NAT).Among 88 patients,95.45%were confirmed as positive by the combination of NAT and antibody test,which was significantly higher(P<0.001)than by single nucleic acid test(73.86%)or serologic test(65.91%).Then the correlation between temporal profile and the level of antibody response was analyzed.It showed that seroconversion started on day 5 after disease onset and IgG level was rose earlier than IgM.Comparison between patients with different disease severity suggested early seroconversion and high antibody titer were linked with less severe clinical symptoms.These results supported the combination of serologic testing and NAT in routine COVID-19 diagnosis and provided evidence on the temporal profile of antibody response in patients with different disease severity.
基金supported by the National Key Research and Development Project of China(2017YFD0201604)the National Natural Science Foundation of China(31571976)。
文摘Potato virus M(PVM) is one of the common and economically important potato viruses in potato-growing regions worldwide. To investigate and control this viral disease, efficient and specific detection techniques are needed. In this study, PVM virions were purified from infected potato plants and used as the immunogen to produce hybridomas secreting PVM-specific monoclonal antibodies(MAbs). Four highly specific and sensitive murine MAbs, i.e., 1 E1, 2 A5, 8 A1 and 17 G8 were prepared through a conventional hybridoma technology. Using these four MAbs, we have developed an antigen-coated plate(ACP)-ELISA, a dot-ELISA and a Tissue print-ELISA for detecting PVM infection in potato plants and tubers. PVM could be detected in infected potato plant tissue crude extracts diluted at 1:10 240(w/v, g mL^(–1)) by the dot-ELISA or at 1:163 840(w/v, g mL^(–1)) by the ACP-ELISA. The Tissue print-ELISA is the quickest and easiest assay among the three established serological assays and is more suitable for onsite large-scale sample detection. Detection results of the field-collected samples showed that PVM is currently widespread in the Yunnan and the Heilongjiang provinces in China. The field sample test results of the developed serological assays were supported by the results from RT-PCR and DNA sequencing. We consider that the newly established ACP-ELISA, dot-ELISA and Tissue print-ELISA can benefit PVM detection in potato plant and tuber samples and field epidemiological studies of PVM. These assays can also facilitate the production of virus-free seed potatoes and breeding for PVM-resistant potato cultivars, leading to the successful prevention of this potato viral disease.
基金Project was supported by the Ministry of Agriculture of China(No.2016ZX08009003-001)the National Key Research and Development Program of China(No.2016YFD0300706)+1 种基金the National Natural Science Foundation of China(No.31571976)the Earmarked Fund for China Agriculture Research System(No.nycytx-001).
文摘Rice stripe mosaic virus(RSMV) is a rhabdovirus recently found in southern part of China and can cause severe reduction in rice production. To establish serological methods for RSMV epidemiological studies and to establish a control strategy for this virus, we first purified RSMV virions from infected rice plants and then used them as an immunogen to produce four RSMV-specific monoclonal antibodies(MAbs)(i.e.,1D4, 4A8, 8E4 and 11F11). With these MAbs, we have developed a highly specific and sensitive antigen-coated plate enzyme-linked immunosorbent assay(ACP-ELISA), a Dot-ELISA and a Tissue print-ELISA for rapid detections of RSMV infection in rice plants or in leafhoppers. Our results showed that RSMV can be readily detected in RSMV-infected rice plant tissue crude extracts diluted at 1:20,971,520(w/v, g/m L)through ACP-ELISA or diluted at 1:327,680(w/v, g/m L) through Dot-ELISA. Both ACP-ELISA and Dot-ELISA can also be used to detect RSMV infection in individual RSMV viruliferous leafhopper(Recilia dorsalis) homogenate diluted at 1:307,200 and 1:163,840(individual leafhopper/l L), respectively. Detection of RSMV infection in field-collected rice samples or in RSMV viruliferous leafhoppers indicated that the three serological methods can produce same results with that produced by RT-PCR(19 of the 33 rice samples and 5 of the 16 leafhoppers were RSMV-positive). We consider that the four MAbs produced in this study are very specific and sensitive, and the three new serological methods are very useful for detections of RSMV infection in rice plants or in leafhoppers and the establishment of the disease control strategies.
文摘BACKGROUND Currently, it is difficult to predict the complications of children at the early stage of sepsis. Brighton pediatric early warning score(PEWS) is a disease risk assessment system that is simple and easy to operate, which has good sensitivity and specificity in disease recognition among children. Because detection indicators vary widely in children, a single indicator is difficult to assess the posttreatment status of children with sepsis.AIM To investigate the relationship between serological markers, Brighton PEWS, and death in children with sepsis after treatment.METHODS A total of 205 children diagnosed with sepsis at our hospital were enrolled. The baseline data, serum scores, and PEWS scores were recorded. In the nested casecontrol study, children who died during the study period were included in an observation group. According to the matching principle, the children who were not dead in the same cohort were included in a control group. The influencing factors of death in children with sepsis after treatment and the value of each evaluation index in predicting the prognosis of children were analyzed.RESULTS A total of 96 children were enrolled in the study, including 48 each in the observation group and the control group. Multivariate logistic regression analysis indicated that antibacterial treatments within 1 h(P = 0.017), shock(P = 0.044),multiple organ dysfunction syndrome(MODS)(P = 0.027), serum procalcitonin(PCT)(P = 0.047), serum albumin(ALB)(P = 0.024), and PEWS(P = 0.012) were independent risk factors for the death of children with sepsis. The area under the curve of the combination of ALB, PCT, and PEWS to predict the death in children with sepsis was the highest(0.908).CONCLUSION Antibacterial treatments within 1 h, shock, MODS, PCT, ALB, and PEWS are independent risk factors for the death of children with sepsis. The predictive accuracy of the combination of PCT, ALB, and PEWS for the prognosis of children with sepsis is the best.
基金Broad Medical Research Program, No. IBD-0119RNIH/NIDDK grant, No. 5R21DK77064+1 种基金NIH/NIDDK, No. KO1-DK62264NIH Ruth L. Kirschstein National Research Service Awards, Proctor & Gamble Investigator Initiated Grants
文摘Serological biomarkers in inflammatory bowel disease (IBD) are a rapidly expanding list of non-invasive tests for objective assessments of disease activity, early diagnosis, prognosis evaluation and surveillance. This review summarizes both old and new biomarkers in IBD, but focuses on the development and character-ization of new serological biomarkers (identifi ed since 2007). These include fi ve new anti-glycan antibodies, anti-chitobioside IgA (ACCA), anti-laminaribioside IgG (ALCA), anti-manobioside IgG (AMCA), and antibod-ies against chemically synthesized (∑) two major oligomannose epitopes, Man α-1,3 Man α-1,2 Man (∑Man3) and Man α-1,3 Man α-1,2 Man α-1,2 Man (∑Man4). These new biomarkers serve as valuable complementary tools to existing biomarkers not only in differentiating Crohn's disease (CD), ulcerative colitis (UC), normal and other non-IBD gut diseases, but also in predicting disease involvement (ileum vs colon), IBD risk (as subclinical biomarkers), and disease course (risk of complication and surgery). Interestingly, the prevalence of the antiglycan antibodies, including anti-Saccharomyces cerevisiae antibodies (ASCA), ALCA and AMCA, was found to be associated with single nucleotide polymorphisms (SNPs) of IBD susceptible genes such as NOD2/CARD15, NOD1/CARD4, toll-likereceptors (TLR) 2 and 4, and β-defensin-1. Further-more, a gene dosage effect was observed: anti-glycan positivity became more frequent as the number of NOD2/CARD15 SNPS increased. Other new serum/ plasma IBD biomarkers reviewed include ubiquitination factor E4A (UBE4A), CXCL16 (a chemokine), resistin, and apolipoprotein A-IV. This review also discusses the most recent studies in IBD biomarker discovery by the application of new technologies such as proteomics, fourier transform near-infrared spectroscopy, and mul-tiplex enzyme-linked immunosorbent assay (ELISA)'s (with an emphasis on cytokine/chemokine profiling). Finally, the prospects of developing more clinically use-ful novel diagnostic algorithms by incorporating new technologies in serological biomarker profiling and integrating multiple biomarkers with bioinformatics analysis/modeling are also discussed.
基金National Key R&D Program of China(Nos.2019YFD1001800 and 2017YFD0201604)the National Natural Science Foundation of China(Nos.31772125 and 31972234)。
文摘Pepino mosaic virus(PepMV)causes severe disease in tomato and other Solanaceous crops around globe.To effectively study and manage this viral disease,researchers need new,sensitive,and high-throughput approaches for viral detection.In this study,we purified PepMV particles from the infected Nicotiana benthamiana plants and used virions to immunize BALB/c mice to prepare hybridomas secreting anti-PepMV monoclonal antibodies(mAbs).A panel of highly specific and sensitive murine mAbs(15B2,8H6,23D11,20D9,3A6,and 8E3)could be produced through cell fusion,antibody selection,and cell cloning.Using the mAbs as the detection antibodies,we established double antibody sandwich enzyme-linked immunosorbent assay(DAS-ELISA),Dot-ELISA,and Tissue print-ELISA for detecting PepMV infection in tomato plants.Resulting data on sensitivity analysis assays showed that both DAS-ELISA and Dot-ELISA can efficiently monitor the virus in PepMV-infected tissue crude extracts when diluted at 1:1310720 and 1:20480(weight/volume ratio(w/v),g/mL),respectively.Among the three methods developed,the Tissue print-ELISA was found to be the most practical detection technique.Survey results from field samples by the established serological approaches were verified by reverse transcription polymerase chain reaction(RT-PCR)and DNA sequencing,dem on strati ng all three serological methods are reliable and effective for monitoring PepMV.An ti-PepMV mAbs and the newly developed DAS-ELISA,Dot-ELISA,and Tissue print-ELISA can benefit PepMV detection and field epidemiological study,and management of this viral disease,which is already widespread in tomato plants in Yunnan Province of China.
基金supported by the National Basic Research Program of China(973 Program)2010CB530200(2010CB530206)the China-US Collaborative Program on Emerging and Re-emerging Infectious Disease(No.1U2GGH000018-01)
文摘Objective:To evaluate the Anaplasma phagocytophilum(A.phagocytophilum),Ehrlichia canis(E.canis,Dirofilaria immitis(D.immitis)(canine heartworm),Borrelia burgdorferi(B.burgdorferi)infections in countryside dogs from Yunnan,Hainan and Anhui provinces.Methods:Serum samples were collected from 26 dogs in Yunnan.Hainan and Anhui provinces.The samples were tested using a commercial ELISA rapid diagnostic assay kit(SNAP^(?)4Dx^(?);IDEXX Laboratories,Inc.U.S.A.).Meaiiwliile,indirect immunofluorescence assay(IFA)recommended by WHO was conducted to delect IgG to A.phagocytophilum.Two methods were analyzed and compared.Results:The number of serologically positive dogs for IgG to A.phagocytophilum was only 2which was from Hainan province and none of the 26 dogs responded positive for E.canu.D.immitis(canine heartworm,and B.burgdorferi by ELISA rapid diagnostic method.The number of serologically positive dogs for IgG to A.phagocytophilum was 13(50%)by IFA method.Data of the two methods were analyzed by statistical software and the difference was statistically significant(P=0.002).Conclusions:It can be concluded that IFA method was more sensitive than ELISA rapid diagnostic method.However,we need conduct further and intensive epidemiology survey on tick-born diseases pathogens including.4.phagocytophilum,E.canis,D.immitis(canine heartworm),and B.burgdorferi which have public health significance.
基金supported by National Institute of Health grants U01 AI061193 and U54-AI057158 (Northeast Biodefense Center).
文摘Many flaviviruses are emerging and reemerging pathogens, such as West Nile virus （WNV）, dengue virus （DENV）, yellow fever virus （YFV）, and Japanese encephalitis virus. Serological assay is the dominant method for diagnosis of flavivirus infections in human. Because antibodies generated during flavivirus infections cross-react with other flavivirus members, plaque reduction neutralization test （PRNT） is the only available assay to determine the infecting flavivirus type. Since PRNT requires culturing raw viruses, it must be performed in biosafety levet-3 or level-4 containment for many flaviviruses, and takes more than ten days to complete. To overcome these problems, we have developed flavivirus viral-like particles （VLPs） that could be used to replace raw viruses in the neutralization assay. The VLPs were prepared by trans packaging a luciferase-reporting replicon with viral structural proteins. This novel assay involves three simple steps： （i） VLPs from a panel of flaviviruses are incubated with flavivirus-infected sera at 37℃ for 1 h; （ii）the neutralized VLPs are used to infect Vero cells; and （iii） the infected cells are measured for luciferase activities at 22 h post-infection. The virus type whose VLP is most efficiently neutralized by the serum specimen （as quantified by the luciferase activities） is the etiologic agent. As a proof-of-concept, we show that a WNV-infected mouse serum neutralized the WNV VLP more efficiently and selectively than the DENV and YFV VLPs. Our results demonstrate that the VLP neutralization assay maintains the ＂gold standard＂ of the classic PRNT; importantly, it shortens the assay time from 〉10 days to 〈1 day, and can be performed in biosafety level-2 facility.
基金supported by National Mega project for Infectious Disease,Ministry of Science and technology(Grant Nos.2016ZX10004222-002,2016ZX10004222-003)National Natural Science Foundation of China(Grant Nos.81373141 and 81401312)National key project of Ebola research,National Natural Science Foundation of China(NSFC,Grant No.81590763)
文摘This study aimed to investigate the serological characteristics of Ebola virus(EBOV) infection during the late phase of the Ebola outbreak in Sierra Leone. In total, 877 blood samples from 694 suspected Ebola virus disease(EVD) cases assessed from March to December 2015, were analyzed via real-time reverse transcription polymerase chain reaction(RT-PCR) for viral RNA and enzyme-linked immunosorbent assay(ELISA) and Luminex to detect antibodies against EBOV. Viral load and EBOV-specific IgM/IgG titers displayed a declining trend during March to December 2015. Viral RNA load decreased rapidly at earlier stages after disease onset, while EBOV-specific IgM and IgG still persisted in 58.1%(18/31) and 93.5%(29/31) of the confirmed EVD patients and in 3.8%(25/663) and 17.8%(118/663) of the RNA-negative suspected patients in the later phase, respectively. Dynamic analysis of longitudinally collected samples from eight EVD patients revealed typically reversed trends of declining viral load and increasing IgM and/or IgG titers in response to the EBOV infection.The present results indicate that certain populations of Sierra Leone developed immunity to an EBOV infection in the late phase of the outbreak, providing novel insights into the risk assessment of EBOV infections among human populations.
基金supported by the National Program on Key Research Project of China(2018YFE0200400,2019YFC1200700)the National Natural Science Foundation of China(U20A20135)+1 种基金the Strategic Biological Resources Capacity Building Project of Chinese Academy of Sciences(KFJ-BRP-017-06)the Key deployment projects of Chinese Academy of Sciences(KJZD-SW-L11)
文摘Severe fever with thrombocytopenia syndrome(SFTS),caused by SFTS virus(SFTSV)infection,was first reported in 2010 in China with an initial fatality of up to 30%.The laboratory confirmation of SFTSV infection in terms of detection of viral RNA or antibody levels is critical for SFTS diagnosis and therapy.In this study,a new luciferase immunoprecipitation system(LIPS)assay based on p REN2 plasmid expressing SFTSV NP gene and tagged with Renilla luciferase(Rluc),was established and used to investigate the levels of antibody responses to SFTSV.Totally 464 serum samples from febrile patients were collected in the hospital of Shaoxing City in Zhejiang Province in 2019.The results showed that 82 of the 464 patients(17.7%)had antibody response to SFTSV,which were further supported by immunofluorescence assays(IFAs).Further,q RT-PCR and microneutralization tests showed that among the 82 positive cases,15 patients had viremia,10 patients had neutralizing antibody,and one had both(totally 26 patient).However,none of these patients were diagnosed as SFTS in the hospital probably because of their mild symptoms or subclinical manifestations.All the results indicated that at least the 26 patients having viremia or neutralizing antibody were the missed diagnosis of SFTS cases.The findings suggested the occurrence of SFTS and the SFTS incidence were higher than the reported level in Shaoxing in 2019,and that LIPS may provide an alternative strategy to confirm SFTSV infection in the laboratory.
基金Supported by The János Bolyai Research and Scholarship of the Hungarian Academy of Sciences,OTKA-K 105530
文摘The spectrum of serological markers associated with inflammatory bowel disease (IBD) is rapidly growing. Due to frequently delayed or missed diagnoses, the application of non-invasive diagnostic tests for IBD, as well as differentiation between ulcerative colitis (UC) and Crohn’s disease (CD), would be useful in the pediatric population. In addition, the combination of pancreatic autoantibodies and antibodies against Saccharomyces cerevisiae antibodies/perinuclear cytoplasmic antibody (pANCA) improved the sensitivity of serological markers in pediatric patients with CD and UC. Some studies suggested that age-associated differences in the patterns of antibodies may be present, particularly in the youngest children. In CD, most patients develop stricturing or perforating complications, and a significant number of patients undergo surgery during the disease course. Based on recent knowledge, serum antibodies are qualitatively and quantitatively associated with complicated CD behavior and CD-related surgery. Pediatric UC is characterized by extensive colitis and a high rate of colectomy. In patients with UC, high levels of anti-CBir1 and pANCA are associated with the development of pouchitis after ileal pouch-anal anastomosis. Thus, serologic markers for IBD can be applied to stratify IBD patients into more homogeneous subgroups with respect to disease progression. In conclusion, identification of patients at an increased risk of rapid disease progression is of great interest, as the application of early and more aggressive pharmaceutical intervention could have the potential to alter the natural history of IBD, and reduce complications and hospitalizations.
