Background Schistosomiasis japonica,caused by Schistosoma japonicum,remains a significant public health concern in the Philippines,where 12.4 million people are at risk due to persistent transmission in endemic region...Background Schistosomiasis japonica,caused by Schistosoma japonicum,remains a significant public health concern in the Philippines,where 12.4 million people are at risk due to persistent transmission in endemic regions.The distribution of schistosomiasis is closely linked to the distribution of its snail intermediate host.This study aims to assess the potential of using soil samples to detect the environmental presence of Oncomelania hupensis quadrasi and Schistosoma japonicum.Methods This cross-sectional observational study utilized a soil-based environmental DNA(eDNA)detection system for simultaneous detection of O.h.quadrasi,and S.japonicum mitochondrially encoded cytochrome c oxidase subunit 1 gene in multiplex quantitative real-time PCR and digital PCR platforms.A two-phase sample collection and testing were carried out in December 2023(Phase 1)and March 2024(Phase 2)across 30 selected sampling sites in Ekiran village,Leyte,Philippines.Wilcoxon two-sample/Mann-Whitney U test was used to determine whether there was a significant difference in eDNA presence and edaphic factors,while percent agreement was used to assess the concordance among methods.Results This study reveals that S.japonicum eDNA can be detected in soil samples and confirms the strong applicability of soil-based eDNA for detecting O.h.quadrasi snails and even in sites without visible snail presence.Furthermore,it demonstrates the superiority of soil-eDNA system compared to classical malacological surveys in detecting O.h.quadrasi and S.japonicum microhabitats:in Phase 1,eDNA detected O.h.quadrasi and S.japonicum in 50%(3/6)and 66.67%(4/6)of sites,respectively,while malacological surveys detected them in only 50%and 16.67%(1/6)of sites.In Phase 2,eDNA detected O.h.quadrasi in 20%(6/30)of sites compared to only 10%(3/30)by malacological survey,and S.japonicum was detected only by eDNA in 10%(3/30)of sites.Among the measured soil parameters,only pH showed a statistically significant difference between eDNA-positive and eDNA-negative sites(P=0.04).Conclusions Soil-based eDNA sensitively detected O.h.quadrasi and S.japonicum,enabling scalable,non-invasive transmission site identification and outperforming traditional surveys without visible snails.Its ability to detect S.japonicum highlights its value for comprehensive schistosomiasis monitoring.展开更多
Background:Schistosomiasis in the People’s Republic of China(PRC)can be traced back to antiquity.In the past 60 years,the Chinese government has made great efforts to control this persistent disease with elimination ...Background:Schistosomiasis in the People’s Republic of China(PRC)can be traced back to antiquity.In the past 60 years,the Chinese government has made great efforts to control this persistent disease with elimination slated by 2020 through the implementation of a comprehensive control strategy.This strategy aims to reduce the role of bovines and humans as sources of infection as a pre-requisite for elimination through transmission interruption.The goal of elimination will be achievable only by the implementation of a sustainable surveillance and control system,with sensitive diagnosis a key feature so that the true disease burden is not underestimated.Currently used diagnostics lack the necessary sensitivity to accurately determine the prevalence of Schistosoma japonicum infection in areas with low infection intensities.It is of critical importance to find and treat people and to identify animals with low-level infections if the National Control Programme for China is to achieve schistosomiasis elimination.Methods:We evaluated a real-time polymerase chain reaction(qPCR)assay using 633 human stool samples collected from five villages in Hunan,Anhui,Hubei,and Jiangxi provinces,and 182 bovine(70 cattle and 112 buffalo)stool samples obtained from four villages in Hunan,Anhui,and Jiangxi provinces in the PRC.All stool samples were subjected to the miracidium hatching test(MHT,a diagnostic procedure used in the National Schistosomiasis Control Programme)and the qPCR assay.Samples positive by MHT were subjected to either the Kato-Katz technique for humans,or the formalin-ethyl acetate sedimentation-digestion(FEA-SD)procedure for bovines,to determine infection intensities.Results:The qPCR assay exhibited a high level of sensitivity in the detection of S.japonicum infections.With both the human and bovine samples,a significantly higher prevalence was determined using the qPCR assay(11.06%humans,24.73%bovines)than with the MHT(0.93%humans,7.69%bovines).The animal contamination index(calculated using data obtained with the qPCR technique)for all positive bovines was 27618000 eggs per day,indicating a considerable amount of environmental egg contamination that would be underestimated using less sensitive diagnostic procedures.Conclusions:The qPCR assay we have evaluated will be applicable as a future field diagnostic and surveillance tool in low-transmission zones where schistosomiasis elimination is targeted and for monitoring post-intervention areas to verify that elimination has been maintained.展开更多
Background:Soil-transmitted helminth(STH)infections have long been an important public health concern in the Philippines.In this review,we describe the current status of STH infections there and highlight the control ...Background:Soil-transmitted helminth(STH)infections have long been an important public health concern in the Philippines.In this review,we describe the current status of STH infections there and highlight the control efforts undertaken to reduce STH burden.Main text:A nationwide STH mass drug administration(MDA)programme was started in 2006 but the overall STH prevalence remains stubbornly high across the Philippines,rangi ng from 24.9%to 97.4%.展开更多
BackgroundSchistosomiasis remains a public health issue and the need for accurate and affordable diagnostics is crucial in the elimination of the disease. While molecular diagnostics are highly effective, they are exp...BackgroundSchistosomiasis remains a public health issue and the need for accurate and affordable diagnostics is crucial in the elimination of the disease. While molecular diagnostics are highly effective, they are expensive, with the main costs been associated with DNA extraction. The DNA dipstick is a rapid, affordable and simple purification method that allows DNA to be extracted from diagnostic samples within 30 s. We aimed to optimise the DNA dipstick method for samples from mice and egg-spiked human samples.MethodsUrine, blood and faeces were collected from mice exposed to Schistosoma japonicum infection at weekly intervals from Day 0 to Day 42. Urine and faecal samples were also collected from volunteer, uninfected humans and spiked with S. japonicum eggs. All samples were subject to several optimisation procedures and DNA extracted with the DNA dipstick. Amplification of the target DNA was carried out using LAMP and visualised using agarose gel electrophoresis and flocculation.ResultsThe DNA dipstick successfully identified S. japonicum from infected mice and human clinical samples spiked with cracked eggs or genomic DNA from S. japonicum. Amplification was observed from week 4 post infection in infected mice. For human samples, amplification was observed in sieved faecal samples, filtered urine samples heated at 95 ℃ for 30 min, and sera samples heated at 95℃ for 30 min.ConclusionsThe DNA dipstick combined with LAMP has huge potential in providing cost-effective, simple and accurate detection of schistosomiasis infection in endemic regions. This will allow for rapid treatment, tracking outbreaks—such as occur after typhoons, leading to better health outcomes and contributing to control and eventual elimination of schistosomiasis.展开更多
基金funded by the e-ASIA Joint Research Program(e-ASIA JRP)Grant No.23jm0210091h0003.
文摘Background Schistosomiasis japonica,caused by Schistosoma japonicum,remains a significant public health concern in the Philippines,where 12.4 million people are at risk due to persistent transmission in endemic regions.The distribution of schistosomiasis is closely linked to the distribution of its snail intermediate host.This study aims to assess the potential of using soil samples to detect the environmental presence of Oncomelania hupensis quadrasi and Schistosoma japonicum.Methods This cross-sectional observational study utilized a soil-based environmental DNA(eDNA)detection system for simultaneous detection of O.h.quadrasi,and S.japonicum mitochondrially encoded cytochrome c oxidase subunit 1 gene in multiplex quantitative real-time PCR and digital PCR platforms.A two-phase sample collection and testing were carried out in December 2023(Phase 1)and March 2024(Phase 2)across 30 selected sampling sites in Ekiran village,Leyte,Philippines.Wilcoxon two-sample/Mann-Whitney U test was used to determine whether there was a significant difference in eDNA presence and edaphic factors,while percent agreement was used to assess the concordance among methods.Results This study reveals that S.japonicum eDNA can be detected in soil samples and confirms the strong applicability of soil-based eDNA for detecting O.h.quadrasi snails and even in sites without visible snail presence.Furthermore,it demonstrates the superiority of soil-eDNA system compared to classical malacological surveys in detecting O.h.quadrasi and S.japonicum microhabitats:in Phase 1,eDNA detected O.h.quadrasi and S.japonicum in 50%(3/6)and 66.67%(4/6)of sites,respectively,while malacological surveys detected them in only 50%and 16.67%(1/6)of sites.In Phase 2,eDNA detected O.h.quadrasi in 20%(6/30)of sites compared to only 10%(3/30)by malacological survey,and S.japonicum was detected only by eDNA in 10%(3/30)of sites.Among the measured soil parameters,only pH showed a statistically significant difference between eDNA-positive and eDNA-negative sites(P=0.04).Conclusions Soil-based eDNA sensitively detected O.h.quadrasi and S.japonicum,enabling scalable,non-invasive transmission site identification and outperforming traditional surveys without visible snails.Its ability to detect S.japonicum highlights its value for comprehensive schistosomiasis monitoring.
基金This work was supported by the Science and Technology Committee of Hunan Province,People’s Republic of China(Grant No.2013WK3023)by the Australian National Health and Medical Research Council(Project Grants:APP1002245,APP1098244,Program Grant:APP1037304).
文摘Background:Schistosomiasis in the People’s Republic of China(PRC)can be traced back to antiquity.In the past 60 years,the Chinese government has made great efforts to control this persistent disease with elimination slated by 2020 through the implementation of a comprehensive control strategy.This strategy aims to reduce the role of bovines and humans as sources of infection as a pre-requisite for elimination through transmission interruption.The goal of elimination will be achievable only by the implementation of a sustainable surveillance and control system,with sensitive diagnosis a key feature so that the true disease burden is not underestimated.Currently used diagnostics lack the necessary sensitivity to accurately determine the prevalence of Schistosoma japonicum infection in areas with low infection intensities.It is of critical importance to find and treat people and to identify animals with low-level infections if the National Control Programme for China is to achieve schistosomiasis elimination.Methods:We evaluated a real-time polymerase chain reaction(qPCR)assay using 633 human stool samples collected from five villages in Hunan,Anhui,Hubei,and Jiangxi provinces,and 182 bovine(70 cattle and 112 buffalo)stool samples obtained from four villages in Hunan,Anhui,and Jiangxi provinces in the PRC.All stool samples were subjected to the miracidium hatching test(MHT,a diagnostic procedure used in the National Schistosomiasis Control Programme)and the qPCR assay.Samples positive by MHT were subjected to either the Kato-Katz technique for humans,or the formalin-ethyl acetate sedimentation-digestion(FEA-SD)procedure for bovines,to determine infection intensities.Results:The qPCR assay exhibited a high level of sensitivity in the detection of S.japonicum infections.With both the human and bovine samples,a significantly higher prevalence was determined using the qPCR assay(11.06%humans,24.73%bovines)than with the MHT(0.93%humans,7.69%bovines).The animal contamination index(calculated using data obtained with the qPCR technique)for all positive bovines was 27618000 eggs per day,indicating a considerable amount of environmental egg contamination that would be underestimated using less sensitive diagnostic procedures.Conclusions:The qPCR assay we have evaluated will be applicable as a future field diagnostic and surveillance tool in low-transmission zones where schistosomiasis elimination is targeted and for monitoring post-intervention areas to verify that elimination has been maintained.
文摘Background:Soil-transmitted helminth(STH)infections have long been an important public health concern in the Philippines.In this review,we describe the current status of STH infections there and highlight the control efforts undertaken to reduce STH burden.Main text:A nationwide STH mass drug administration(MDA)programme was started in 2006 but the overall STH prevalence remains stubbornly high across the Philippines,rangi ng from 24.9%to 97.4%.
文摘BackgroundSchistosomiasis remains a public health issue and the need for accurate and affordable diagnostics is crucial in the elimination of the disease. While molecular diagnostics are highly effective, they are expensive, with the main costs been associated with DNA extraction. The DNA dipstick is a rapid, affordable and simple purification method that allows DNA to be extracted from diagnostic samples within 30 s. We aimed to optimise the DNA dipstick method for samples from mice and egg-spiked human samples.MethodsUrine, blood and faeces were collected from mice exposed to Schistosoma japonicum infection at weekly intervals from Day 0 to Day 42. Urine and faecal samples were also collected from volunteer, uninfected humans and spiked with S. japonicum eggs. All samples were subject to several optimisation procedures and DNA extracted with the DNA dipstick. Amplification of the target DNA was carried out using LAMP and visualised using agarose gel electrophoresis and flocculation.ResultsThe DNA dipstick successfully identified S. japonicum from infected mice and human clinical samples spiked with cracked eggs or genomic DNA from S. japonicum. Amplification was observed from week 4 post infection in infected mice. For human samples, amplification was observed in sieved faecal samples, filtered urine samples heated at 95 ℃ for 30 min, and sera samples heated at 95℃ for 30 min.ConclusionsThe DNA dipstick combined with LAMP has huge potential in providing cost-effective, simple and accurate detection of schistosomiasis infection in endemic regions. This will allow for rapid treatment, tracking outbreaks—such as occur after typhoons, leading to better health outcomes and contributing to control and eventual elimination of schistosomiasis.
