Colorectal cancer(CRC)is one of the most prevalent cancers in developed countries.On the other hand,CRC is also one of the most curable cancers if it is detected in early stages through regular colonoscopy or sigmoido...Colorectal cancer(CRC)is one of the most prevalent cancers in developed countries.On the other hand,CRC is also one of the most curable cancers if it is detected in early stages through regular colonoscopy or sigmoidoscopy.Since CRC develops slowly from precancerous lesions,early detection can reduce both the incidence and mortality of the disease.Fecal occult blood test is a widely used non-invasive screening tool for CRC.Although fecal occult blood test is simple and cost-effective in screening CRC,there is room for improvement in terms of the accuracy of the test.Genetic dysregulations have been found to play an important role in CRC development.With better understanding of the molecular basis of CRC,there is a growing expectation on the development of diagnostic tests based on more sensitive and specific molecular markers and those tests may provide a breakthrough to the limitations of current screening tests for CRC.In this review,the molecular basis of CRC development,the characteristics and applications of different non-invasive molecular biomarkers,as well as the technologies available for the detection were discussed.This review intended to provide a summary on the current and future molecular diagnostics in CRC and its pre-malignant state,colorectal adenoma.展开更多
Traditional diagnostic strategies for infectious disease detection require benchtop instruments that are inappropriate for point-of-care testing(POCT). Emerging microfluidics, a highly miniaturized, automatic, and int...Traditional diagnostic strategies for infectious disease detection require benchtop instruments that are inappropriate for point-of-care testing(POCT). Emerging microfluidics, a highly miniaturized, automatic, and integrated technology,are a potential substitute for traditional methods in performing rapid, low-cost, accurate, and on-site diagnoses.Molecular diagnostics are widely used in microfluidic devices as the most effective approaches for pathogen detection.This review summarizes the latest advances in microfluidics-based molecular diagnostics for infectious diseases from academic perspectives and industrial outlooks. First, we introduce the typical on-chip nucleic acid processes,including sample preprocessing, amplification, and signal read-out. Then, four categories of microfluidic platforms are compared with respect to features, merits, and demerits. We further discuss application of the digital assay in absolute nucleic acid quantification. Both the classic and recent microfluidics-based commercial molecular diagnostic devices are summarized as proof of the current market status. Finally, we propose future directions for microfluidics-based infectious disease diagnosis.展开更多
Morphology,immunology,cytogenetics,and molecular biology(MICM)diagnostic systems have become the industry standard in the fields of hematological disease diagnosis and treatment.1 With the rapid development of molecul...Morphology,immunology,cytogenetics,and molecular biology(MICM)diagnostic systems have become the industry standard in the fields of hematological disease diagnosis and treatment.1 With the rapid development of molecular biology and genomics technologies,the molecular diagnosis and treatment of blood tumors have made remarkable progress,providing patients with more precise and effective treatment strategies.The special issue,which is titled“Precision medicine in hematological cancers”,is focused on numerous challenges in the treatment of hematological malignancies,including drug resistance,adverse events,and disease recurrence,and explore methods to integrate the latest molecular assays to improve diagnostic accuracy and treatment safety.展开更多
Along with the advances in cancer genomics and the development of targeted therapies, the field of molecular diagnostics has undergone rapid evolution to meet the growing needs associated with patient care. Here, we r...Along with the advances in cancer genomics and the development of targeted therapies, the field of molecular diagnostics has undergone rapid evolution to meet the growing needs associated with patient care. Here, we review the past, present, and possible future of molecular diagnostics, including technologies and testing principles, to provide a comprehensive landscape of molecular diagnostic technologies, testing platforms, and applications. This review is based on the US Food and Drug Administration publications, the National Comprehensive Cancer Network guidelines, and the peer-reviewed English literature published between 2003 and 2021. We conclude that molecular diagnostics has changed dramatically during the past two decades. Next-generation sequencing–based comprehensive genomic profiling has replaced single-gene/single-locus testing for simultaneous detection of mutations, copy number alterations, structural variants, and mutational signatures to facilitate cancer diagnosis, prognosis prediction, targeted therapies, and immunotherapies. Laboratory-developed tests and companion diagnostics approved by the US Food and Drug Administration both play important roles in cancer patient management.展开更多
Precise tumor diagnosis and treatment require the support of abundant molecular information.However,conventional molecular diagnostic technologies gradually fail to satisfy the demands of clinical therapy due to limit...Precise tumor diagnosis and treatment require the support of abundant molecular information.However,conventional molecular diagnostic technologies gradually fail to satisfy the demands of clinical therapy due to limited detection performance.Benefiting from highly specific target sequence recognition and efficient cis/trans cleavage activity,CRISPR/Cas system has been widely employed to construct novel molecular diagnostic strategies,hailed as the“next-generation molecular diagnostic technology”.This review focuses on recent advances in CRISPR molecular diagnostic systems for the detection of tumor variant gene,protein,and liquid biopsy biomarker,and outlines strategies for CRISPR in situ molecular detection.In addition,we explore general principles and development trends in the construction of CRISPR molecular diagnostic system and emphasize the revolutionary impact that it has brought to the field of molecular diagnostics.展开更多
Uterine tumors resembling ovarian sex cord tumors(UTROSCTs)are characterized by an uncertain malignant potential and exhibit prominent sex cord-like differentiation.The purpose of this study was to comprehensively rev...Uterine tumors resembling ovarian sex cord tumors(UTROSCTs)are characterized by an uncertain malignant potential and exhibit prominent sex cord-like differentiation.The purpose of this study was to comprehensively review the clinicopathological characteristics of UTROSCTs and analyze eight cases of UTROSCTs treated at our hospital.We conducted an extensive review of the relevant literature and gathered pertinent data.In addition,we identified eight patients with UTROSCTs and analyzed their clinical and pathological features,diagnosis,treatment,and prognosis.Patients presented with symptoms such as abnormal vaginal bleeding or uterine mass detection.Surgical interventions varied,including total abdominal hysterectomy,bilateral salpingo-oophorectomy,and pelvic lymphadenectomy,with adjuvant therapy given to one patient.All eight patients are currently disease-free,with the longest follow-up period being nearly 10 years.Our systematic review of UTROSCTs summarized the clinical and pathological features and revealed several novel markers,including ESR1-NCOA2-3,GREB1-NCOA1-3,GREB1-CTNNB1,and GREB1-NR4A3.UTROSCTs are rare mesenchymal tumors with unclear histogenesis and uncertain malignant potential.Although our understanding of UTROSCTs remains incomplete,the promising findings and increasing availability of clinical data will contribute to the further understanding and development of this rare neoplasm.展开更多
In Burkina Faso, as in other African countries, infertility has become a social burden for the population and a public health problem. Male infertility accounts for 30% to 40% of all infertility cases. The diagnosis o...In Burkina Faso, as in other African countries, infertility has become a social burden for the population and a public health problem. Male infertility accounts for 30% to 40% of all infertility cases. The diagnosis of male infertility or hypofertility is often made by a simple laboratory analysis of sperm to explore sperm parameters. In most African countries, such as Burkina Faso, microbiological analysis in the context of sperm analysis is still not developed, and is carried out solely based on microscopy and traditional culture, which does not allow the growth of fragile and demanding bacteria. Our study investigated the microorganisms of sperm that may be involved in male infertility, using conventional bacteriology techniques and real-time PCR. However, it did not intend to perform a multivariate statistical association analysis to estimate the association of microorganisms with abnormal semen parameters. This prospective cross-sectional pilot study was carried out on patients who visited the bacteriology laboratory of Centre MURAZ, a research Institute in Burkina Faso, for male infertility diagnosis between 2 August and 31 August 2021. Bacteria were isolated and identified using standard bacteriology techniques. In parallel, common pathogenic microorganisms known to be associated with male infertility were targeted and detected in the sperm using a multiplex real-time PCR assay. A total of 38 sperm samples were analyzed by bacteriological culture and bacteria isolated were Staphylococcus aureus (S. aureus) 5.55%, Klebsiella pneumoniae (K. pneumoniae), Enterococcus faecalis (E. faecalis), Streptococcus agalactiae (S. agalactiae) and Staphylococcus hoemalyticus (S. hoemalyticus) respectively 2.70%. Real-time PCR targeted and detected Chlamydia trachomatis (C. trachomatis) at 7.89%, Ureaplasma urealyticum (U. urealyticum) at 21.05%, Ureaplasma parvum (U. parvum) at 18.42%, Mycoplasma hominis (M. hominis) at 15.79%, Mycoplasma genitalium (M. genitalium) at 10.53% and Trichomonas vaginalis (T. vaginalis) at 2.63%. Neisseria gonorrhoeae (N. gonorrhoeae) was targeted by the real-time PCR assay and was not detected (0%) in the tested semen samples. Our study highlights critical limitations of culture performance (low sensitivity), particularly in Burkina Faso, which has a total inability to detect microorganisms (fragile and demanding microorganisms) detected by PCR-based assays. There is therefore an urgent need to at least optimize culture, procedures and algorithms for detection of microorganisms associated with male infertility in clinical laboratories of Burkina Faso. The most effective solution is the routine implementation of molecular diagnostic methods.展开更多
Sexually transmitted infections (STIs) represent a public health problem due to their high prevalence worldwide and the emergence of multidrug resistance of responsible microorganisms. Medical laboratory diagnosis of ...Sexually transmitted infections (STIs) represent a public health problem due to their high prevalence worldwide and the emergence of multidrug resistance of responsible microorganisms. Medical laboratory diagnosis of sexually transmitted genital infections by traditional methods as culture remains extremely delicate, difficult or impossible (to find extremely fragile organisms that can be cultured). Thus, molecular techniques constitute an alternative to improve accurate diagnostic, personalized patient treatment, and public health. A total of 83 clinical samples including urethral discharge and urine samples from individual patients with symptoms of urethritis received were analyzed using traditional methods and a commercial real-time PCR (qPCR) method. Out of 83 urethritis patients, n = 55 (66.26%) were positive for at least one of the STI pathogens detected by qPCR. qPCR assay was more sensitive (50/83, positive cases) compared to culture (15/83, positive cases) and light microscopy (28/83, positive cases). The most prevalent NTD pathogen in the suspected patients was N. gonorrhoeae with 60.24% (50/83) based on real-time PCR diagnosis. Among the positive cases of STI pathogens, Neisseria gonorrhoeae had the highest frequency 49/55 (89.01%) followed by low frequencies of Trichomonas vaginalis 4/55 (7.27%) and Chlamydia trachomatis 1/55 (1.82%). This highlights the high prevalence of N. gonorrhoeae infection in male urethritis patients and a very important misdiagnosis using traditional routine methods in Burkina Faso by medical laboratories. Thus, this situation may negatively impact patients’ personalized treatment and care and public health with the possible rapid emergence of multidrug-resistant strains. This study also highlights the urgent need to optimize culture for the diagnosis of NTD pathogens in Burkina Faso and the usefulness and the need for the introduction of molecular diagnostic methods in routine diagnosis for the detection of NTD pathogens in the medical laboratories in Burkina Faso.展开更多
Objective:To establish Nipah virus diagnostic capabilities at the National Reference Laboratory in Sri Lanka using the NIV Pune real-time PCR kit.Methods:Strict safety precautions were adhered during testing due to th...Objective:To establish Nipah virus diagnostic capabilities at the National Reference Laboratory in Sri Lanka using the NIV Pune real-time PCR kit.Methods:Strict safety precautions were adhered during testing due to the high pathogenicity of the Nipah virus,with all diagnostics conducted in a BSL2+laboratory at the Medical Research Institute in Sri Lanka.RNA extraction was performed using the QIAamp Viral RNA Mini kit.The NIV Pune in-house real-time PCR kit was employed,following established primer/probe sequences and controls.The assay was validated using the Rotor-Gene Q Series Real-time PCR platform.Results:The validation run of the Nipah virus real-time PCR test demonstrated robust performance,with positive controls consistently detecting Nipah RNA at a Ct value of 21.50±0.01.Negative controls confirmed assay specificity with an external negative control which was also used as an extraction control and showed no interference.The internal control exhibited stable behavior,enhancing confidence in PCR results.The qPCR analysis graph illustrated the successful detection of internal and positive controls,validating the reliability of the assay.Conclusions:Establishing Nipah virus diagnostic capabilities in Sri Lanka signifies a proactive and collaborative response to the persistent global health threat.展开更多
Molecular diagnostics is one of the most important tools currently in use for clinical pathogen detection due to its high sensitivity,specificity,and low consume of sample and reagent is keyword to low cost molecular ...Molecular diagnostics is one of the most important tools currently in use for clinical pathogen detection due to its high sensitivity,specificity,and low consume of sample and reagent is keyword to low cost molecular diagnostics.In this paper,a sensitive DNA isothermal amplifi-cation method for fast clinical infectious diseases diagnostics at aM concentrations of DNA was developed using a polycarbonate(PC)microfuidic chip.A portable confocal optical fuo-rescence detector was specifically developed for the microfuidic chip that was capable of highly sensitive real-time detection of amplified products for sequence-specific molecular identification near the optical diffraction limit with low background.The molecular diagnostics of Listeria monocytogenes with nucleic acid extracted from stool samples was performed at a minimum DNA template concentration of 3.65 aM,and a detection limit of less than five copies of genomic DNA.Contrast to the general polymerase chain reaction(PCR)at eppendorf(EP)tube,the detection time in our developed method was reduced from 1.5h to 45 min for multi-target parallel detection,the consume of sample and reagent was dropped from 25μL to 1.45μL.This novel microfuidic chip system and method can be used to develop a micro total analysis system as a clinically relevant pathogen molecular diagnostics method via the amplification of targets,with potential applications in biotechnology,medicine,and clinical molecular diagnostics.展开更多
DNA sequencing is the method of identifying the precise order of DNA nucleotides within a molecule. The information of DNA sequencing is of prime requisite for basic biological research as well as in various clinical ...DNA sequencing is the method of identifying the precise order of DNA nucleotides within a molecule. The information of DNA sequencing is of prime requisite for basic biological research as well as in various clinical specialties.They can be used to determine the individual genetic sequence, larger genetic regions, chromosomes as well as to sequence RNA and proteins. Since the first DNA sequencing in 1970s, there has been tremendous advancements in the technologies aimed to determine the entire human genome. The need for rapid and accurate sequencing of human genome has resulted in the introduction of next generation sequencing(NGS) technology. NGS refers to the secondgeneration DNA sequencing technologies where millions of DNA can be sequenced simultaneously. Some of the next gen sequencing methods employed are Roche/454 life science, Illumina/Solexa, SOLiD system and HeliScope.Application of NGS in decoding the genomic database of various oral diseases may possess therapeutic and prognostic value. This presentation provides an overview of the basics of NGS and their potential applications in oral disease diagnostics.展开更多
To apply an individualized oncological approach to gastric cancer patients,the accurate diagnosis of disease entities is required.Peritoneal metastasis is the most frequent mode of metastasis in gastric cancer,and the...To apply an individualized oncological approach to gastric cancer patients,the accurate diagnosis of disease entities is required.Peritoneal metastasis is the most frequent mode of metastasis in gastric cancer,and the tumor-node-metastasis classification includes cytological detection of intraperitoneal cancer cells as part of the staging process,denoting metastatic disease.The accuracy of cytological diagnosis leaves room for improvement;therefore,highly sensitive molecular diagnostics,such as an enzyme immunoassay,reverse transcription polymerase chain reaction,and virusguided imaging,have been developed to detect minute cancer cells in the peritoneal cavity.Molecular targeting therapy has also been spun off from basic research in the past decade.Although conventional cytologyis still the mainstay,novel approaches could serve as practical complementary diagnostics to cytology in near future.展开更多
Dear Editor：Quantitative real-time PCR has revolutionized molecular diagnostics with its ease of use,increased sensitivity and specificity and low turnaround time.PCR/quantitative PCR（qPCR）-based assays offer a dis...Dear Editor：Quantitative real-time PCR has revolutionized molecular diagnostics with its ease of use,increased sensitivity and specificity and low turnaround time.PCR/quantitative PCR（qPCR）-based assays offer a distinct advantage over other serological/conventional diagnostic approaches.The ability to diagnose infectious diseases has benefited from the availability of US FDA approved and Conformite Europeenne（CE）-marked qPCR-based in-vitro diagnostic kits from international companies.The high-quality kits are calibrated with the World Health Organization（WHO）reference standards and the National Institute for Biological Standards and Control（NIBSC）standards.展开更多
Pancreatic cystic lesions are being increasingly detected,mainly due to the widespread use of cross-sectional imaging.The reported prevalence ranges from 13%to 18%in asymptomatic individuals.These lesions display a br...Pancreatic cystic lesions are being increasingly detected,mainly due to the widespread use of cross-sectional imaging.The reported prevalence ranges from 13%to 18%in asymptomatic individuals.These lesions display a broad histologic spectrum,from benign pseudocysts to premalignant mucinous cystic neoplasms and invasive carcinomas.Although many classification and management strategies exist,the natural history of numerous pancreatic cystic lesions remains incompletely understood,contributing to significant clinical uncertainty.Current diagnostic tools,including computed tomography,magnetic resonance imaging,endoscopic ultrasound,and cyst fluid analysis,are constrained by either suboptimal sensitivity or high costs.Cytology,while specific when positive,suffers from low sensitivity.Biochemical markers such as carcinoembryonic antigen,amylase,and glucose can help in cyst differentiation,whereas molecular testing(e.g.,KRAS,GNAS,RNF43 mutations)provides additional diagnostic and prognostic value.However,the application of molecular diagnostics is still restricted in routine practice due to costs,access issues,and a lack of standardization.This diagnostic uncertainty leads to both overtreatment and undertreatment.Some patients undergo unnecessary surgeries for benign lesions,which exposes them to procedural risks and long-term consequences.Others may experience delays in interventions for high-risk cysts and missing opportunities for cancer prevention.Additionally,prolonged,and often unnecessary surveillance burdens patients and healthcare systems psychologically and financially.In this minireview,we present a comprehensive overview of the classification,diagnostic approach,and management of pancreatic cystic lesions,incorporating recent evidence and current international guidelines(Fukuoka,American Gastroenterological Association,European).We also highlight the limitations of existing strategies and emerging tools such as radiomics,next-generation sequencing,and novel biomarkers.Additionally,we emphasize the urgent need for cost-effective,accurate,and accessible diagnostic pathways.A more refined risk stratification approach is essential to optimize outcomes,reduce healthcare waste,and improve the quality of life for patients with pancreatic cystic lesions.展开更多
Granulicatella species,previously classified as nutritionally variant streptococci,have emerged as important opportunistic pathogens.Although typically part of the normal oral,gastrointestinal,and genitourinary flora,...Granulicatella species,previously classified as nutritionally variant streptococci,have emerged as important opportunistic pathogens.Although typically part of the normal oral,gastrointestinal,and genitourinary flora,these fastidious Gram-positive cocci can cause serious infections in both immunocompetent and immunocompromised individuals.Their identification often proves difficult due to slow growth and unusual nutritional requirements,which limit the reliability of conventional culture methods.This narrative review consolidates data from over 100 case reports and clinical studies to present a comprehensive overview of Granulicatella infections.Infective endocarditis appears as the most frequently reported and life-threatening condition,often affecting the aortic and mitral valves.Patients with prosthetic valves or congenital heart defects face a higher risk.Delayed diagnosis frequently results in complications such as heart failure,systemic emboli,and renal dysfunction.Granulicatella species also contribute to osteomyelitis,pulmonary infections,brain abscesses,gastrointestinal sepsis,and infections related to implanted medical devices.Effective management typically involves beta-lactam antibiotics combined with aminoglycosides,although antimicrobial resistance remains a concern.Surgical intervention often proves necessary,especially in cases with prosthetic material or severe complications.Outcomes are generally favorable when infections are identified early and managed effectively.However,delayed recognition and suboptimal therapy continue to contribute to poor prognoses in some patients.This review emphasizes the need to improve clinical awareness and establish standardized diagnostic and therapeutic approaches to address the challenges posed by Granulicatella infections.展开更多
Dermatofibrosarcoma protuberans(DFSP)is a rare cutaneous intermediate-grade soft tissue tumor characterized by COL1A1::PDGFB fusion in most cases.This fusion drives tumorigenesis and forms the basis for imatinib treat...Dermatofibrosarcoma protuberans(DFSP)is a rare cutaneous intermediate-grade soft tissue tumor characterized by COL1A1::PDGFB fusion in most cases.This fusion drives tumorigenesis and forms the basis for imatinib treatment,which acts by blocking platelet-derived growth factor receptor-beta kinase activity.Apart from this canonical fusion,there is an expanding spectrum of rare fusions,including COL6A3::PDGFD,EMILIN::PDGFD,TNC::PDGFD,etc.,through mole-cular profiling.These atypical rearrangements may be encountered in morpho-logically classic DFSP,unusual anatomic sites,or diagnostically challenging variants such as fibrosarcomatous DFSP.Their recognition is clinically relevant,as they may influence tumor biology,response to targeted therapy,and eligibility for clinical trials.This newly documented DFSP involving the lacrimal sac was initially misdiagnosed as a solitary fibrous tumor,emphasizing the diagnostic pitfalls in anatomically constrained regions and the importance of integrated diagnosis combining histology,immunohistochemistry,and molecular testing.In this editorial commentary,we briefly highlight the ever-growing genomic land-scape of DFSP,report rare fusions and their biological implications,and examine the role of expanded molecular diagnostics in refining diagnosis,guiding therapy,and informing prognosis.Incorporating comprehensive fusion analysis into routine workup may be critical for accurate classification,especially in unusual presentations where reliance on morphology alone risks misdiagnosis.展开更多
AIM: To determine the frequency of occult hepatitis B infection (OHBI) in a group of human immunodeficiency virus (HIV)-1+/ hepatitis B surface antigen negative (HBsAg)- patients from Mexico.
Human papillomavirus(HPV)infection identified as a definitive human carcinogen is increasingly being recognized for its role in carcinogenesis of human cancers.Up to 38%–80%of head and neck squamous cell carcinoma(HN...Human papillomavirus(HPV)infection identified as a definitive human carcinogen is increasingly being recognized for its role in carcinogenesis of human cancers.Up to 38%–80%of head and neck squamous cell carcinoma(HNSCC)in oropharyngeal location(OPSCC)and nearly all cervical cancers contain the HPV genome which is implicated in causing cancer through its oncoproteins E6 and E7.Given by the biologically distinct HPV-related OPSCC and a more favorable prognosis compared to HPV-negative tumors,clinical trials on de-escalation treatment strategies for these patients have been studied.It is therefore raised the questions for the patient stratification if treatment de-escalation is feasible.Moreover,understanding the crosstalk of HPV-mediated malignancy and immunity with clinical insights from the proportional response rate to immune checkpoint blockade treatments in patients with HNSCC is of importance to substantially improve the treatment efficacy.This review discusses the biology of HPV-related HNSCC as well as successful clinically findings with promising candidates in the pipeline for future directions.With the advent of various sequencing technologies,further biomolecules associated with HPV-related HNSCC progression are currently being identified to be used as potential biomarkers or targets for clinical decisions throughout the continuum of cancer care.展开更多
AIM: To estimate the prevalence of human immunodeficiency virus (HIV) and hepatitis C virus (HCV) infections in women in Mali and to evaluate the performance of serological assays.
The Zika virus(ZIKV) is an arbovirus that has spread rapidly worldwide within recent times. There is accumulating evidence that associates ZIKV infections with Guillain-Barré Syndrome(GBS) and microcephaly in hum...The Zika virus(ZIKV) is an arbovirus that has spread rapidly worldwide within recent times. There is accumulating evidence that associates ZIKV infections with Guillain-Barré Syndrome(GBS) and microcephaly in humans. The ZIKV is genetically diverse and can be separated into Asian and African lineages. A rapid, sensitive, and specific assay is needed for the detection of ZIKV across various pandemic regions. So far, the available primers and probes do not cover the genetic diversity and geographic distribution of all ZIKV strains. To this end, we have developed a one-step quantitative reverse transcription polymerase chain reaction(qRT-PCR) assay based on conserved sequences in the ZIKV envelope(E) gene. The detection limit of the assay was determined to be five RNA transcript copies and 2.94 × 10^(–3) 50% tissue culture infectious doses(TCID50) of live ZIKV per reaction. The assay was highly specific and able to detect five different ZIKV strains covering the Asian and African lineages without nonspecific amplification, when tested against other flaviviruses. The assay was also successful in testing for ZIKV in clinical samples. Our assay represents an improvement over the current methods available for the detection ZIKV and would be valuable as a diagnostic tool in various pandemic regions.展开更多
文摘Colorectal cancer(CRC)is one of the most prevalent cancers in developed countries.On the other hand,CRC is also one of the most curable cancers if it is detected in early stages through regular colonoscopy or sigmoidoscopy.Since CRC develops slowly from precancerous lesions,early detection can reduce both the incidence and mortality of the disease.Fecal occult blood test is a widely used non-invasive screening tool for CRC.Although fecal occult blood test is simple and cost-effective in screening CRC,there is room for improvement in terms of the accuracy of the test.Genetic dysregulations have been found to play an important role in CRC development.With better understanding of the molecular basis of CRC,there is a growing expectation on the development of diagnostic tests based on more sensitive and specific molecular markers and those tests may provide a breakthrough to the limitations of current screening tests for CRC.In this review,the molecular basis of CRC development,the characteristics and applications of different non-invasive molecular biomarkers,as well as the technologies available for the detection were discussed.This review intended to provide a summary on the current and future molecular diagnostics in CRC and its pre-malignant state,colorectal adenoma.
基金supported by the Nationa l Key Research and Development Program of China (2021YFA1101500)the National Natural Science Foundation of China (22074047)+1 种基金the Hubei Provincial Natural Science Foundation of China (2020CFB578)the Fundamental Research Funds for Central Universities,HUST (2020kfy XJJS034)。
文摘Traditional diagnostic strategies for infectious disease detection require benchtop instruments that are inappropriate for point-of-care testing(POCT). Emerging microfluidics, a highly miniaturized, automatic, and integrated technology,are a potential substitute for traditional methods in performing rapid, low-cost, accurate, and on-site diagnoses.Molecular diagnostics are widely used in microfluidic devices as the most effective approaches for pathogen detection.This review summarizes the latest advances in microfluidics-based molecular diagnostics for infectious diseases from academic perspectives and industrial outlooks. First, we introduce the typical on-chip nucleic acid processes,including sample preprocessing, amplification, and signal read-out. Then, four categories of microfluidic platforms are compared with respect to features, merits, and demerits. We further discuss application of the digital assay in absolute nucleic acid quantification. Both the classic and recent microfluidics-based commercial molecular diagnostic devices are summarized as proof of the current market status. Finally, we propose future directions for microfluidics-based infectious disease diagnosis.
文摘Morphology,immunology,cytogenetics,and molecular biology(MICM)diagnostic systems have become the industry standard in the fields of hematological disease diagnosis and treatment.1 With the rapid development of molecular biology and genomics technologies,the molecular diagnosis and treatment of blood tumors have made remarkable progress,providing patients with more precise and effective treatment strategies.The special issue,which is titled“Precision medicine in hematological cancers”,is focused on numerous challenges in the treatment of hematological malignancies,including drug resistance,adverse events,and disease recurrence,and explore methods to integrate the latest molecular assays to improve diagnostic accuracy and treatment safety.
文摘Along with the advances in cancer genomics and the development of targeted therapies, the field of molecular diagnostics has undergone rapid evolution to meet the growing needs associated with patient care. Here, we review the past, present, and possible future of molecular diagnostics, including technologies and testing principles, to provide a comprehensive landscape of molecular diagnostic technologies, testing platforms, and applications. This review is based on the US Food and Drug Administration publications, the National Comprehensive Cancer Network guidelines, and the peer-reviewed English literature published between 2003 and 2021. We conclude that molecular diagnostics has changed dramatically during the past two decades. Next-generation sequencing–based comprehensive genomic profiling has replaced single-gene/single-locus testing for simultaneous detection of mutations, copy number alterations, structural variants, and mutational signatures to facilitate cancer diagnosis, prognosis prediction, targeted therapies, and immunotherapies. Laboratory-developed tests and companion diagnostics approved by the US Food and Drug Administration both play important roles in cancer patient management.
基金supported by the National Natural Science Foundation of China(Nos.82330030 and 82121003)the Sichuan Science and Technology Program(Nos.2021YFS0369 and 2024NSFSC1541)the CAMS Innovation Fund for Medical Sciences(No.2019-12M-5-032).
文摘Precise tumor diagnosis and treatment require the support of abundant molecular information.However,conventional molecular diagnostic technologies gradually fail to satisfy the demands of clinical therapy due to limited detection performance.Benefiting from highly specific target sequence recognition and efficient cis/trans cleavage activity,CRISPR/Cas system has been widely employed to construct novel molecular diagnostic strategies,hailed as the“next-generation molecular diagnostic technology”.This review focuses on recent advances in CRISPR molecular diagnostic systems for the detection of tumor variant gene,protein,and liquid biopsy biomarker,and outlines strategies for CRISPR in situ molecular detection.In addition,we explore general principles and development trends in the construction of CRISPR molecular diagnostic system and emphasize the revolutionary impact that it has brought to the field of molecular diagnostics.
文摘Uterine tumors resembling ovarian sex cord tumors(UTROSCTs)are characterized by an uncertain malignant potential and exhibit prominent sex cord-like differentiation.The purpose of this study was to comprehensively review the clinicopathological characteristics of UTROSCTs and analyze eight cases of UTROSCTs treated at our hospital.We conducted an extensive review of the relevant literature and gathered pertinent data.In addition,we identified eight patients with UTROSCTs and analyzed their clinical and pathological features,diagnosis,treatment,and prognosis.Patients presented with symptoms such as abnormal vaginal bleeding or uterine mass detection.Surgical interventions varied,including total abdominal hysterectomy,bilateral salpingo-oophorectomy,and pelvic lymphadenectomy,with adjuvant therapy given to one patient.All eight patients are currently disease-free,with the longest follow-up period being nearly 10 years.Our systematic review of UTROSCTs summarized the clinical and pathological features and revealed several novel markers,including ESR1-NCOA2-3,GREB1-NCOA1-3,GREB1-CTNNB1,and GREB1-NR4A3.UTROSCTs are rare mesenchymal tumors with unclear histogenesis and uncertain malignant potential.Although our understanding of UTROSCTs remains incomplete,the promising findings and increasing availability of clinical data will contribute to the further understanding and development of this rare neoplasm.
文摘In Burkina Faso, as in other African countries, infertility has become a social burden for the population and a public health problem. Male infertility accounts for 30% to 40% of all infertility cases. The diagnosis of male infertility or hypofertility is often made by a simple laboratory analysis of sperm to explore sperm parameters. In most African countries, such as Burkina Faso, microbiological analysis in the context of sperm analysis is still not developed, and is carried out solely based on microscopy and traditional culture, which does not allow the growth of fragile and demanding bacteria. Our study investigated the microorganisms of sperm that may be involved in male infertility, using conventional bacteriology techniques and real-time PCR. However, it did not intend to perform a multivariate statistical association analysis to estimate the association of microorganisms with abnormal semen parameters. This prospective cross-sectional pilot study was carried out on patients who visited the bacteriology laboratory of Centre MURAZ, a research Institute in Burkina Faso, for male infertility diagnosis between 2 August and 31 August 2021. Bacteria were isolated and identified using standard bacteriology techniques. In parallel, common pathogenic microorganisms known to be associated with male infertility were targeted and detected in the sperm using a multiplex real-time PCR assay. A total of 38 sperm samples were analyzed by bacteriological culture and bacteria isolated were Staphylococcus aureus (S. aureus) 5.55%, Klebsiella pneumoniae (K. pneumoniae), Enterococcus faecalis (E. faecalis), Streptococcus agalactiae (S. agalactiae) and Staphylococcus hoemalyticus (S. hoemalyticus) respectively 2.70%. Real-time PCR targeted and detected Chlamydia trachomatis (C. trachomatis) at 7.89%, Ureaplasma urealyticum (U. urealyticum) at 21.05%, Ureaplasma parvum (U. parvum) at 18.42%, Mycoplasma hominis (M. hominis) at 15.79%, Mycoplasma genitalium (M. genitalium) at 10.53% and Trichomonas vaginalis (T. vaginalis) at 2.63%. Neisseria gonorrhoeae (N. gonorrhoeae) was targeted by the real-time PCR assay and was not detected (0%) in the tested semen samples. Our study highlights critical limitations of culture performance (low sensitivity), particularly in Burkina Faso, which has a total inability to detect microorganisms (fragile and demanding microorganisms) detected by PCR-based assays. There is therefore an urgent need to at least optimize culture, procedures and algorithms for detection of microorganisms associated with male infertility in clinical laboratories of Burkina Faso. The most effective solution is the routine implementation of molecular diagnostic methods.
文摘Sexually transmitted infections (STIs) represent a public health problem due to their high prevalence worldwide and the emergence of multidrug resistance of responsible microorganisms. Medical laboratory diagnosis of sexually transmitted genital infections by traditional methods as culture remains extremely delicate, difficult or impossible (to find extremely fragile organisms that can be cultured). Thus, molecular techniques constitute an alternative to improve accurate diagnostic, personalized patient treatment, and public health. A total of 83 clinical samples including urethral discharge and urine samples from individual patients with symptoms of urethritis received were analyzed using traditional methods and a commercial real-time PCR (qPCR) method. Out of 83 urethritis patients, n = 55 (66.26%) were positive for at least one of the STI pathogens detected by qPCR. qPCR assay was more sensitive (50/83, positive cases) compared to culture (15/83, positive cases) and light microscopy (28/83, positive cases). The most prevalent NTD pathogen in the suspected patients was N. gonorrhoeae with 60.24% (50/83) based on real-time PCR diagnosis. Among the positive cases of STI pathogens, Neisseria gonorrhoeae had the highest frequency 49/55 (89.01%) followed by low frequencies of Trichomonas vaginalis 4/55 (7.27%) and Chlamydia trachomatis 1/55 (1.82%). This highlights the high prevalence of N. gonorrhoeae infection in male urethritis patients and a very important misdiagnosis using traditional routine methods in Burkina Faso by medical laboratories. Thus, this situation may negatively impact patients’ personalized treatment and care and public health with the possible rapid emergence of multidrug-resistant strains. This study also highlights the urgent need to optimize culture for the diagnosis of NTD pathogens in Burkina Faso and the usefulness and the need for the introduction of molecular diagnostic methods in routine diagnosis for the detection of NTD pathogens in the medical laboratories in Burkina Faso.
文摘Objective:To establish Nipah virus diagnostic capabilities at the National Reference Laboratory in Sri Lanka using the NIV Pune real-time PCR kit.Methods:Strict safety precautions were adhered during testing due to the high pathogenicity of the Nipah virus,with all diagnostics conducted in a BSL2+laboratory at the Medical Research Institute in Sri Lanka.RNA extraction was performed using the QIAamp Viral RNA Mini kit.The NIV Pune in-house real-time PCR kit was employed,following established primer/probe sequences and controls.The assay was validated using the Rotor-Gene Q Series Real-time PCR platform.Results:The validation run of the Nipah virus real-time PCR test demonstrated robust performance,with positive controls consistently detecting Nipah RNA at a Ct value of 21.50±0.01.Negative controls confirmed assay specificity with an external negative control which was also used as an extraction control and showed no interference.The internal control exhibited stable behavior,enhancing confidence in PCR results.The qPCR analysis graph illustrated the successful detection of internal and positive controls,validating the reliability of the assay.Conclusions:Establishing Nipah virus diagnostic capabilities in Sri Lanka signifies a proactive and collaborative response to the persistent global health threat.
基金the National Natural Science Foundation of China(81327005,61361160418,61575100)the National Foundation of High Technology of China(2012 AA020102,2013 AA041201)+2 种基金the National Key Foundation for Exploring Scientific Instruments(2013 YQ190467)the Beijing Municipal Natural Science Foundation(4142025)the Beijing Lab Foundation,and the Tsinghua Autonomous Research Foundation(2014 Z01001)。
文摘Molecular diagnostics is one of the most important tools currently in use for clinical pathogen detection due to its high sensitivity,specificity,and low consume of sample and reagent is keyword to low cost molecular diagnostics.In this paper,a sensitive DNA isothermal amplifi-cation method for fast clinical infectious diseases diagnostics at aM concentrations of DNA was developed using a polycarbonate(PC)microfuidic chip.A portable confocal optical fuo-rescence detector was specifically developed for the microfuidic chip that was capable of highly sensitive real-time detection of amplified products for sequence-specific molecular identification near the optical diffraction limit with low background.The molecular diagnostics of Listeria monocytogenes with nucleic acid extracted from stool samples was performed at a minimum DNA template concentration of 3.65 aM,and a detection limit of less than five copies of genomic DNA.Contrast to the general polymerase chain reaction(PCR)at eppendorf(EP)tube,the detection time in our developed method was reduced from 1.5h to 45 min for multi-target parallel detection,the consume of sample and reagent was dropped from 25μL to 1.45μL.This novel microfuidic chip system and method can be used to develop a micro total analysis system as a clinically relevant pathogen molecular diagnostics method via the amplification of targets,with potential applications in biotechnology,medicine,and clinical molecular diagnostics.
文摘DNA sequencing is the method of identifying the precise order of DNA nucleotides within a molecule. The information of DNA sequencing is of prime requisite for basic biological research as well as in various clinical specialties.They can be used to determine the individual genetic sequence, larger genetic regions, chromosomes as well as to sequence RNA and proteins. Since the first DNA sequencing in 1970s, there has been tremendous advancements in the technologies aimed to determine the entire human genome. The need for rapid and accurate sequencing of human genome has resulted in the introduction of next generation sequencing(NGS) technology. NGS refers to the secondgeneration DNA sequencing technologies where millions of DNA can be sequenced simultaneously. Some of the next gen sequencing methods employed are Roche/454 life science, Illumina/Solexa, SOLiD system and HeliScope.Application of NGS in decoding the genomic database of various oral diseases may possess therapeutic and prognostic value. This presentation provides an overview of the basics of NGS and their potential applications in oral disease diagnostics.
基金Supported by Grants from Ministry of Education,Science,and Culture,Japan(to Kagawa S)and JSPS KAKENHI,No.23591932
文摘To apply an individualized oncological approach to gastric cancer patients,the accurate diagnosis of disease entities is required.Peritoneal metastasis is the most frequent mode of metastasis in gastric cancer,and the tumor-node-metastasis classification includes cytological detection of intraperitoneal cancer cells as part of the staging process,denoting metastatic disease.The accuracy of cytological diagnosis leaves room for improvement;therefore,highly sensitive molecular diagnostics,such as an enzyme immunoassay,reverse transcription polymerase chain reaction,and virusguided imaging,have been developed to detect minute cancer cells in the peritoneal cavity.Molecular targeting therapy has also been spun off from basic research in the past decade.Although conventional cytologyis still the mainstay,novel approaches could serve as practical complementary diagnostics to cytology in near future.
文摘Dear Editor：Quantitative real-time PCR has revolutionized molecular diagnostics with its ease of use,increased sensitivity and specificity and low turnaround time.PCR/quantitative PCR（qPCR）-based assays offer a distinct advantage over other serological/conventional diagnostic approaches.The ability to diagnose infectious diseases has benefited from the availability of US FDA approved and Conformite Europeenne（CE）-marked qPCR-based in-vitro diagnostic kits from international companies.The high-quality kits are calibrated with the World Health Organization（WHO）reference standards and the National Institute for Biological Standards and Control（NIBSC）standards.
文摘Pancreatic cystic lesions are being increasingly detected,mainly due to the widespread use of cross-sectional imaging.The reported prevalence ranges from 13%to 18%in asymptomatic individuals.These lesions display a broad histologic spectrum,from benign pseudocysts to premalignant mucinous cystic neoplasms and invasive carcinomas.Although many classification and management strategies exist,the natural history of numerous pancreatic cystic lesions remains incompletely understood,contributing to significant clinical uncertainty.Current diagnostic tools,including computed tomography,magnetic resonance imaging,endoscopic ultrasound,and cyst fluid analysis,are constrained by either suboptimal sensitivity or high costs.Cytology,while specific when positive,suffers from low sensitivity.Biochemical markers such as carcinoembryonic antigen,amylase,and glucose can help in cyst differentiation,whereas molecular testing(e.g.,KRAS,GNAS,RNF43 mutations)provides additional diagnostic and prognostic value.However,the application of molecular diagnostics is still restricted in routine practice due to costs,access issues,and a lack of standardization.This diagnostic uncertainty leads to both overtreatment and undertreatment.Some patients undergo unnecessary surgeries for benign lesions,which exposes them to procedural risks and long-term consequences.Others may experience delays in interventions for high-risk cysts and missing opportunities for cancer prevention.Additionally,prolonged,and often unnecessary surveillance burdens patients and healthcare systems psychologically and financially.In this minireview,we present a comprehensive overview of the classification,diagnostic approach,and management of pancreatic cystic lesions,incorporating recent evidence and current international guidelines(Fukuoka,American Gastroenterological Association,European).We also highlight the limitations of existing strategies and emerging tools such as radiomics,next-generation sequencing,and novel biomarkers.Additionally,we emphasize the urgent need for cost-effective,accurate,and accessible diagnostic pathways.A more refined risk stratification approach is essential to optimize outcomes,reduce healthcare waste,and improve the quality of life for patients with pancreatic cystic lesions.
文摘Granulicatella species,previously classified as nutritionally variant streptococci,have emerged as important opportunistic pathogens.Although typically part of the normal oral,gastrointestinal,and genitourinary flora,these fastidious Gram-positive cocci can cause serious infections in both immunocompetent and immunocompromised individuals.Their identification often proves difficult due to slow growth and unusual nutritional requirements,which limit the reliability of conventional culture methods.This narrative review consolidates data from over 100 case reports and clinical studies to present a comprehensive overview of Granulicatella infections.Infective endocarditis appears as the most frequently reported and life-threatening condition,often affecting the aortic and mitral valves.Patients with prosthetic valves or congenital heart defects face a higher risk.Delayed diagnosis frequently results in complications such as heart failure,systemic emboli,and renal dysfunction.Granulicatella species also contribute to osteomyelitis,pulmonary infections,brain abscesses,gastrointestinal sepsis,and infections related to implanted medical devices.Effective management typically involves beta-lactam antibiotics combined with aminoglycosides,although antimicrobial resistance remains a concern.Surgical intervention often proves necessary,especially in cases with prosthetic material or severe complications.Outcomes are generally favorable when infections are identified early and managed effectively.However,delayed recognition and suboptimal therapy continue to contribute to poor prognoses in some patients.This review emphasizes the need to improve clinical awareness and establish standardized diagnostic and therapeutic approaches to address the challenges posed by Granulicatella infections.
文摘Dermatofibrosarcoma protuberans(DFSP)is a rare cutaneous intermediate-grade soft tissue tumor characterized by COL1A1::PDGFB fusion in most cases.This fusion drives tumorigenesis and forms the basis for imatinib treatment,which acts by blocking platelet-derived growth factor receptor-beta kinase activity.Apart from this canonical fusion,there is an expanding spectrum of rare fusions,including COL6A3::PDGFD,EMILIN::PDGFD,TNC::PDGFD,etc.,through mole-cular profiling.These atypical rearrangements may be encountered in morpho-logically classic DFSP,unusual anatomic sites,or diagnostically challenging variants such as fibrosarcomatous DFSP.Their recognition is clinically relevant,as they may influence tumor biology,response to targeted therapy,and eligibility for clinical trials.This newly documented DFSP involving the lacrimal sac was initially misdiagnosed as a solitary fibrous tumor,emphasizing the diagnostic pitfalls in anatomically constrained regions and the importance of integrated diagnosis combining histology,immunohistochemistry,and molecular testing.In this editorial commentary,we briefly highlight the ever-growing genomic land-scape of DFSP,report rare fusions and their biological implications,and examine the role of expanded molecular diagnostics in refining diagnosis,guiding therapy,and informing prognosis.Incorporating comprehensive fusion analysis into routine workup may be critical for accurate classification,especially in unusual presentations where reliance on morphology alone risks misdiagnosis.
基金Supported by Consejo Nacional de Ciencia y Tecnologia,Mexico CONACYT 2008-C01-86717,(to Alvarez-Muoz MT and Lira R)
文摘AIM: To determine the frequency of occult hepatitis B infection (OHBI) in a group of human immunodeficiency virus (HIV)-1+/ hepatitis B surface antigen negative (HBsAg)- patients from Mexico.
基金This work was supported by grants from the Medical and the Health Science Project of Zhejiang Province(2019KY327)Guangji Talents Foundation Award(E)of Zhejiang Cancer Hospital。
文摘Human papillomavirus(HPV)infection identified as a definitive human carcinogen is increasingly being recognized for its role in carcinogenesis of human cancers.Up to 38%–80%of head and neck squamous cell carcinoma(HNSCC)in oropharyngeal location(OPSCC)and nearly all cervical cancers contain the HPV genome which is implicated in causing cancer through its oncoproteins E6 and E7.Given by the biologically distinct HPV-related OPSCC and a more favorable prognosis compared to HPV-negative tumors,clinical trials on de-escalation treatment strategies for these patients have been studied.It is therefore raised the questions for the patient stratification if treatment de-escalation is feasible.Moreover,understanding the crosstalk of HPV-mediated malignancy and immunity with clinical insights from the proportional response rate to immune checkpoint blockade treatments in patients with HNSCC is of importance to substantially improve the treatment efficacy.This review discusses the biology of HPV-related HNSCC as well as successful clinically findings with promising candidates in the pipeline for future directions.With the advent of various sequencing technologies,further biomolecules associated with HPV-related HNSCC progression are currently being identified to be used as potential biomarkers or targets for clinical decisions throughout the continuum of cancer care.
文摘AIM: To estimate the prevalence of human immunodeficiency virus (HIV) and hepatitis C virus (HCV) infections in women in Mali and to evaluate the performance of serological assays.
基金supported by the National Science and Technology Major Project(2016ZX10004222)the Sanming Project of Medicine in Shenzhen(ZDSYS201504301534057)+6 种基金the Key specialized fund for infectious diseases in Shenzhen City(No.201161)the intramural special grant for influenza virus research from the Chinese Academy of Sciences(KJZD-EW-L09 and KJZD-EWL15)the Shenzhen Science and Technology Research and Development Project(JCYJ20160427151920801 and JCYJ20160427153238750)G.F.G.is a leading principal investigator of the National Natural Science Foundation of China(NSFC)Innovative Research Group(81621091)supported by the Youth Innovation Promotion Association of Chinese Academy of Sciences(CAS)(2017122)G.W.is the recipient of a Banting Postdoctoral Fellowship from the Canadian Institutes of Health Research(CIHR)the President’s International Fellowship Initiative from the CAS
文摘The Zika virus(ZIKV) is an arbovirus that has spread rapidly worldwide within recent times. There is accumulating evidence that associates ZIKV infections with Guillain-Barré Syndrome(GBS) and microcephaly in humans. The ZIKV is genetically diverse and can be separated into Asian and African lineages. A rapid, sensitive, and specific assay is needed for the detection of ZIKV across various pandemic regions. So far, the available primers and probes do not cover the genetic diversity and geographic distribution of all ZIKV strains. To this end, we have developed a one-step quantitative reverse transcription polymerase chain reaction(qRT-PCR) assay based on conserved sequences in the ZIKV envelope(E) gene. The detection limit of the assay was determined to be five RNA transcript copies and 2.94 × 10^(–3) 50% tissue culture infectious doses(TCID50) of live ZIKV per reaction. The assay was highly specific and able to detect five different ZIKV strains covering the Asian and African lineages without nonspecific amplification, when tested against other flaviviruses. The assay was also successful in testing for ZIKV in clinical samples. Our assay represents an improvement over the current methods available for the detection ZIKV and would be valuable as a diagnostic tool in various pandemic regions.
