RESUMO
BACKGROUND: Schistosomiasis japonica represents a significant public health concern in South Asia. There is an urgent need to optimize existing schistosomiasis diagnostic techniques. This study aims to develop models for the different stages of liver fibrosis caused by Schistosoma infection utilizing ultrasound radiomics and machine learning techniques. METHODS: From 2018 to 2022, we retrospectively collected data on 1,531 patients and 5,671 B-mode ultrasound images from the Second People's Hospital of Duchang City, Jiangxi Province, China. The datasets were screened based on inclusion and exclusion criteria suitable for radiomics models. Liver fibrosis due to Schistosoma infection (LFSI) was categorized into four stages: grade 0, grade 1, grade 2, and grade 3. The data were divided into six binary classification problems, such as group 1 (grade 0 vs. grade 1) and group 2 (grade 0 vs. grade 2). Key radiomic features were extracted using Pyradiomics, the Mann-Whitney U test, and the Least Absolute Shrinkage and Selection Operator (LASSO). Machine learning models were constructed using Support Vector Machine (SVM), and the contribution of different features in the model was described by applying Shapley Additive Explanations (SHAP). RESULTS: This study ultimately included 1,388 patients and their corresponding images. A total of 851 radiomics features were extracted for each binary classification problems. Following feature selection, 18 to 76 features were retained from each groups. The area under the receiver operating characteristic curve (AUC) for the validation cohorts was 0.834 (95% CI: 0.779-0.885) for the LFSI grade 0 vs. LFSI grade 1, 0.771 (95% CI: 0.713-0.835) for LFSI grade 1 vs. LFSI grade 2, and 0.830 (95% CI: 0.762-0.885) for LFSI grade 2 vs. LFSI grade 3. CONCLUSION: Machine learning models based on ultrasound radiomics are feasible for classifying different stages of liver fibrosis caused by Schistosoma infection.
Assuntos
Estudos de Viabilidade , Cirrose Hepática , Schistosoma japonicum , Esquistossomose Japônica , Ultrassonografia , Humanos , Esquistossomose Japônica/diagnóstico por imagem , Ultrassonografia/métodos , Masculino , Cirrose Hepática/diagnóstico por imagem , Feminino , Estudos Retrospectivos , Pessoa de Meia-Idade , Adulto , Schistosoma japonicum/classificação , Schistosoma japonicum/isolamento & purificação , China , Animais , Aprendizado de Máquina , Máquina de Vetores de Suporte , Idoso , Adulto Jovem , Adolescente , Fígado/diagnóstico por imagem , Fígado/parasitologia , Fígado/patologia , RadiômicaRESUMO
Schistosoma species are the causative agent of schistosomiasis and shows worldwide distribution. There is a great need to develop a sensitive diagnostic approach for controlling the disease. Previously, we identified large numbers of Extracellular Vesicle (EV) proteins from Schistosoma japonicum (S. japonicum), but rarely these proteins have been evaluated for their diagnostic potential. In the present study, we performed bioinformatic analyses of S. japonicum identified EV-associated proteins from the previous study and then identified Schistosoma-specific proteins with potentially secreted capability. Among them, we selected SJCHGC02838 protein, SJCHGC05593 protein, SJCHGC05668 protein and a hypothetical protein (SJHYP) to evaluate their diagnostic potential for detecting S. japonicum infection. First, we determined the expression of these four proteins at the transcript levels using qRT-PCR and revealed that all these genes showed higher expression in adult stage. Then, we cloned the full-length cDNA for each protein into a prokaryotic expression vector and successfully generated the recombinant proteins. Upon the purification of recombinant proteins, we developed an indirect ELISA method to evaluate the diagnostic potential of these purified recombinant proteins. The results showed high sensitivity for detecting Schistosoma infection. Additionally, these proteins also displayed a good potential for detecting Schistosoma infection, especially SJCHGC05668 protein at an early stage. The diagnostic potentials of these recombinant proteins were further evaluated by Western blot and comparatively analyzed by our previously developed cfDNA methods.
Assuntos
Biomarcadores , Ensaio de Imunoadsorção Enzimática , Vesículas Extracelulares , Proteínas de Helminto , Schistosoma japonicum , Esquistossomose Japônica , Schistosoma japonicum/genética , Schistosoma japonicum/metabolismo , Schistosoma japonicum/isolamento & purificação , Animais , Vesículas Extracelulares/metabolismo , Vesículas Extracelulares/genética , Esquistossomose Japônica/diagnóstico , Esquistossomose Japônica/parasitologia , Proteínas de Helminto/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/isolamento & purificação , Biologia Computacional , Sensibilidade e Especificidade , Camundongos , Humanos , Feminino , Clonagem MolecularRESUMO
BACKGROUND: Schistosomiasis is a neglected tropical parasitic and chronic disease affecting hundreds of millions of people. Adult schistosomes reside in the blood stream of the definitive mammalian host. These helminth parasites possess two epithelial surfaces, the tegument and the gastrodermis, both of which interact with the host during immune evasion and in nutrient uptake. METHODS: Female ARC Swiss mice (4-6 weeks old) were infected percutaneously with Schistosoma japonicum cercariae freshly shed from Oncomelania hupensis quadrasi snails (Philippines strain). Fluorescent in situ hybridisation (FISH) was performed by using fresh adult S. japonicum perfused from those infected mice. Adult S. japonicum worms were processed to isolate the tegument from the carcass containing the gastrodermis; blood and bile were collected individually from infected and uninfected mice. Total DNA extracted from all those samples were used for microbiome profiling. RESULTS: FISH and microbiome profiling showed the presence of bacterial populations on two epithelial surfaces of adult worms, suggesting they were distinct not only from the host blood but also from each other. Whereas microbial diversity was reduced overall in the parasite epithelial tissues when compared with that of host blood, specific bacterial taxa, including Anoxybacillus and Escherichia, were elevated on the tegument. Minimal differences were evident in the microbiome of host blood during an active infection, compared with that of control uninfected blood. However, sampling of bile from infected animals identified some differences compared with controls, including elevated levels of Limnohabitans, Clostridium and Curvibacter. CONCLUSIONS: Using FISH and microbial profiling, we were able to demonstrate, for the first time, that bacteria are presented on the epithelial surfaces of adult schistosomes. These schistosome surface-associated bacteria, which are distinct from the host blood microenvironment, should be considered as a new and important component of the host-schistosome interaction. The importance of individual bacterial species in relation to schistosome parasitism needs further elucidation.
Assuntos
Sangue/microbiologia , Epitélio/microbiologia , Microbiota/genética , Schistosoma japonicum/microbiologia , Esquistossomose Japônica/sangue , Animais , Anoxybacillus/genética , Bile/microbiologia , Cercárias , Clostridium/genética , Comamonadaceae/genética , DNA Bacteriano/genética , DNA Bacteriano/isolamento & purificação , Modelos Animais de Doenças , Escherichia coli/genética , Feminino , Hibridização in Situ Fluorescente/métodos , Masculino , Camundongos , RNA Ribossômico 16S/genética , Schistosoma japonicum/isolamento & purificação , Esquistossomose Japônica/parasitologia , Caramujos/parasitologiaRESUMO
BACKGROUND: Elimination and control of Schistosoma japonicum, the most virulent of the schistosomiasis-causing blood flukes, requires the development of sensitive and specific diagnostic tools capable of providing an accurate measurement of the infection prevalence in endemic areas. Typically, detection of S. japonicum has occurred using the Kato-Katz technique, but this methodology, which requires skilled microscopists, has been shown to radically underestimate levels of infection. With the ever-improving capabilities of next-generation sequencing and bioinformatic analysis tools, identification of satellite sequences and other highly repetitive genomic elements for use as real-time PCR diagnostic targets is becoming increasingly common. Assays developed using these targets have the ability to improve the sensitivity and specificity of results for epidemiological studies that can in turn be used to inform mass drug administration and programmatic decision making. METHODOLOGY/PRINCIPAL FINDINGS: Utilizing Tandem Repeat Analyzer (TAREAN) and RepeatExplorer2, a cluster-based analysis of the S. japonicum genome was performed and a tandemly arranged genomic repeat, which we named SjTR1 (Schistosoma japonicum Tandem Repeat 1), was selected as the target for a real-time PCR diagnostic assay. Based on these analyses, a primer/probe set was designed and the assay was optimized. The resulting real-time PCR test was shown to reliably detect as little as 200 ag of S. japonicum genomic DNA and as little as 1 egg per gram of human stool. Based on these results, the index assay reported in this manuscript is more sensitive than previously published real-time PCR assays for the detection of S. japonicum. CONCLUSIONS/SIGNIFICANCE: The extremely sensitive and specific diagnostic assay described in this manuscript will facilitate the accurate detection of S. japonicum, particularly in regions with low levels of endemicity. This assay will be useful in providing data to inform programmatic decision makers, aiding disease control and elimination efforts.
Assuntos
Fezes/parasitologia , Reação em Cadeia da Polimerase em Tempo Real/métodos , Schistosoma japonicum/isolamento & purificação , Esquistossomose Japônica/parasitologia , Animais , Primers do DNA/genética , Feminino , Humanos , Masculino , Schistosoma japonicum/genética , Esquistossomose Japônica/diagnóstico , Sensibilidade e EspecificidadeRESUMO
BACKGROUND: Zoonotic schistosomiasis, caused by Schistosoma japonicum, remains a major public health problem in the Philippines. This study aimed to evaluate the commercially available rapid diagnostic point-of-care circulating cathodic antigen (POC-CCA) test in detecting individuals infected with S. japonicum in a human cohort from an endemic area for schistosomiasis japonica in the Philippines. METHODS: Clinical samples were collectedin 18 barangays endemic for S. japonicum infection in Laoang and Palapag municipalities, Northern Samar, the Philippines, in 2015. The presence of CCA in filter-concentrated urine samples (n = 412) was evaluated using the commercial kits and the results were converted to images, which were further analyzed by ImageJ software to calculate R values. The diagnostic performance of the immunochromatographic POC-CCA test was compared using the Kato-Katz (KK) procedure, in-house enzyme-linked immunosorbent assays (ELISAs) and droplet digital (dd) PCR assays as reference. RESULTS: The POC-CCA test was able to detect S. japonicum-infected individuals in the cohort with an eggs per gram of faeces (EPG) more than or equal to 10 with sensitivity/specificity values of 63.3%/93.3%. However, the assay showed an inability to diagnose schistosomiasis japonica infections in all cohort KK-positive individuals, of which the majority had an extremely low egg burden (EPG: 1-9). The prevalence of S. japonicum infection in the total cohort determined by the POC-CCA test was 12.4%, only half of that determined by the KK method (26.2%). When compared with the ELISAs and ddPCR assays as a reference, the POC-CCA assay was further shown to be a test with low sensitivity. Nevertheless, the assay exhibited significant positive correlations with egg burden determined by the KK technique and the target gene copy number index values determined by the ddPCR assays within the entire cohort. CONCLUSIONS: By using in silico image analysis, the POC-CCA cassette test could be converted to a quantitative assay to avoid reader-variability. Because of its low sensitivity, the commercially available POC-CCA assay had limited potential for determining the status of a S. japonicum infection in the target cohort. The assay should be applied with caution in populations where schistosome parasites (especially S. japonicum) are present at low infection intensity.
Assuntos
Antígenos de Helmintos/sangue , Fezes/parasitologia , Sistemas Automatizados de Assistência Junto ao Leito , Schistosoma japonicum/isolamento & purificação , Esquistossomose Japônica/diagnóstico , Testes Sorológicos/métodos , Animais , Anti-Helmínticos/uso terapêutico , Antígenos de Helmintos/análise , Ensaio de Imunoadsorção Enzimática , Humanos , Doenças Negligenciadas/epidemiologia , Filipinas/epidemiologia , Reação em Cadeia da Polimerase , Prevalência , Schistosoma japonicum/imunologia , Esquistossomose Japônica/tratamento farmacológico , Esquistossomose Japônica/epidemiologia , Sensibilidade e EspecificidadeRESUMO
BACKGROUND: Oncomelania hupensis is only intermediate snail host of Schistosoma japonicum, and distribution of O. hupensis is an important indicator for the surveillance of schistosomiasis. This study explored the feasibility of a random forest algorithm weighted by spatial distance for risk prediction of schistosomiasis distribution in the Yangtze River Basin in China, with the aim to produce an improved precision reference for the national schistosomiasis control programme by reducing the number of snail survey sites without losing predictive accuracy. METHODS: The snail presence and absence records were collected from Anhui, Hunan, Hubei, Jiangxi and Jiangsu provinces in 2018. A machine learning of random forest algorithm based on a set of environmental and climatic variables was developed to predict the breeding sites of the O. hupensis intermediated snail host of S. japonicum. Different spatial sizes of a hexagonal grid system were compared to estimate the need for required snail sampling sites. The predictive accuracy related to geographic distances between snail sampling sites was estimated by calculating Kappa and the area under the curve (AUC). RESULTS: The highest accuracy (AUC = 0.889 and Kappa = 0.618) was achieved at the 5 km distance weight. The five factors with the strongest correlation to O. hupensis infestation probability were: (1) distance to lake (48.9%), (2) distance to river (36.6%), (3) isothermality (29.5%), (4) mean daily difference in temperature (28.1%), and (5) altitude (26.0%). The risk map showed that areas characterized by snail infestation were mainly located along the Yangtze River, with the highest probability in the dividing, slow-flowing river arms in the middle and lower reaches of the Yangtze River in Anhui, followed by areas near the shores of China's two main lakes, the Dongting Lake in Hunan and Hubei and the Poyang Lake in Jiangxi. CONCLUSIONS: Applying the machine learning of random forest algorithm made it feasible to precisely predict snail infestation probability, an approach that could improve the sensitivity of the Chinese schistosome surveillance system. Redesign of the snail surveillance system by spatial bias correction of O. hupensis infestation in the Yangtze River Basin to reduce the number of sites required to investigate from 2369 to 1747.
Assuntos
Rios/parasitologia , Schistosoma japonicum/isolamento & purificação , Caramujos/fisiologia , Animais , Cruzamento , China , Monitoramento Ambiental , Estudos de Viabilidade , Aprendizado de Máquina , Modelos Teóricos , Vigilância da População , Caramujos/parasitologia , Análise EspacialRESUMO
OBJECTIVE: Schistosomiasis japonica is an important helminthic disease in Asia. Sensitive and accurate diagnostic tools are indispensable for clinical diagnosis, screening infection and monitoring its control. In this study, we developed an immunochromatographic test (Sj-ICT) to detect anti-Schistosoma japonicum immunoglobulin G antibodies in human sera. METHODS: Somatic extract from adult S. japonicum was used as an antigen. The Sj-ICT was developed and optimized as a point-of-care test. All 214 human serum samples were evaluated for diagnostic usefulness and comparison with an enzyme-linked immunosorbent assay (ELISA). RESULTS: The diagnostic sensitivity, specificity, positive and negative predictive values, and accuracy of the Sj-ICT were 90.8%, 87.9%, 86.4%, 91.9% and 89.3%, respectively. For ELISA the values were respectively 91.8%, 87.9%, 86.5%, 92.7% and 89.7%. The concordance between both methods was 86.4 % (Cohen's kappa value = 0.729). CONCLUSIONS: The immunochromatographic test kit developed can support clinical diagnosis and large-scale surveys in endemic areas without requiring additional facilities or ancillary supplies.
Assuntos
Anticorpos Anti-Helmínticos/sangue , Ensaio de Imunoadsorção Enzimática/métodos , Imunoensaio/métodos , Imunoglobulina G/sangue , Testes Imediatos , Esquistossomose Japônica/diagnóstico , Animais , Ásia , Feminino , Humanos , Masculino , Valor Preditivo dos Testes , Schistosoma japonicum/imunologia , Schistosoma japonicum/isolamento & purificação , Sensibilidade e EspecificidadeRESUMO
Molecular diagnostics are powerful tools for disease detection but are typically confined to the laboratory environment due to the cumbersome methods required to extract nucleic acids from biological samples. Accurate diagnosis is essential for early detection of parasitic worm infections and for monitoring control programs, particularly during new transmission outbreaks to limit infection spread. We optimized the recently developed DNA dipstick technology to purify Schistosoma japonicum DNA from different life stages in <60 s. We successfully detected DNA from adult worms, eggs and infected snails. The speed and simplicity of this method enables the point-of-care detection of S. japonicum.
Assuntos
DNA de Helmintos/isolamento & purificação , Schistosoma japonicum/isolamento & purificação , Esquistossomose Japônica/diagnóstico , Animais , Fígado/parasitologia , Camundongos , Técnicas de Diagnóstico Molecular , Técnicas de Amplificação de Ácido Nucleico , Testes Imediatos , Reação em Cadeia da Polimerase em Tempo Real , Schistosoma japonicum/genética , Esquistossomose Japônica/prevenção & controle , Caramujos/parasitologiaRESUMO
Schistosomiasis persists in Asian regions despite aggressive elimination measures. To identify factors enabling continued parasite transmission, we performed reduced representation genome sequencing on Schistosoma japonicum miracidia collected across multiple years from transmission hotspots in Sichuan, China. We discovered strong geographic structure, suggesting that local, rather than imported, reservoirs are key sources of persistent infections in the region. At the village level, parasites collected after referral for praziquantel treatment are closely related to local pre-treatment populations. Schistosomes within villages are also highly related, suggesting that only a few parasites from a limited number of hosts drive re-infection. The close familial relationships among miracidia from different human hosts also implicate short transmission routes among humans. At the individual host level, genetic evidence indicates that multiple humans retained infections following referral for treatment. Our findings suggest that end-game schistosomiasis control measures should focus on completely extirpating local parasite reservoirs and confirming successful treatment of infected human hosts.
Assuntos
Variação Genética , Metagenômica , Schistosoma japonicum/genética , Esquistossomose Japônica/parasitologia , Seleção Genética , Animais , China/epidemiologia , Genótipo , Humanos , Schistosoma japonicum/classificação , Schistosoma japonicum/isolamento & purificação , Esquistossomose Japônica/epidemiologia , Esquistossomose Japônica/transmissãoRESUMO
BACKGROUND: Most of national schistosomiasis elimination programmes in Asia are relying on stool examination, particularly Kato Katz stool examination technique for regular transmission monitoring. However, the Kato-Katz technique has shown low sensitivity for the detection of light-intensity infections, and therefore highly sensitive diagnostic tools are urgently required to monitor prevalence of infection in low transmission settings. The objective of this systematic review was to evaluate and synthesize the performance of diagnostic tests for detecting Schistosoma japonicum and S. mekongi infection in people living in endemic areas. METHODOLOGY/PRINCIPAL FINDINGS: We comprehensively searched these nine electronic databases and other resources until July 2019, with no language or publication limits: PubMed, EMBASE, MEDLINE, Web of Science, BIOSIS Citation Index, HTA, CINAHL PLUS, The Cochrane Library, and PsycINFO. We included original studies that assessed diagnostic performance using antibody, antigen, and molecular tests with stool examination test as a reference standard. Two reviewers independently extracted a standard set of data and assessed study quality. We estimated the pooled estimates of sensitivity and specificity for each index test. We used diagnostic odds ratio to determine the overall accuracy and hierarchical summary receiver operating characteristics (HSROC) curve to assess the index tests performance. Fifteen studies (S. japonicum [n = 13] and S. mekongi [n = 2]) testing 15,303 participants were included in the review. Five studies reported performance of enzyme-linked immunosorbent assay (ELISA), seven studies reported indirect hemagglutination assay (IHA), and four studies reported polymerase chain reaction (PCR) for detecting S. japonicum. The pooled sensitivity and specificity were 0.93 (95% CI: 0.84-0.98) and 0.40 (95% CI: 0.29-0.53) for ELISA, 0.97 (95% CI: 0.90-0.99) and 0.66 (95% CI: 0.58-0.73) for IHA, and 0.89 (95% CI: 0.71-0.96) and 0.49 (95% CI: 0.29-0.69) for PCR respectively. A global summary indicated the best performance for IHA, closely followed by ELISA. We were unable to perform meta-analysis for S. mekongi due to insufficient number of studies. CONCLUSIONS/SIGNIFICANCE: IHA showed the highest detection accuracy for S. japonicum. Further studies are needed to determine the suitable diagnostic methods to verify the absence of transmission of S. mekongi and also to compare detection accuracy against more sensitive reference standards such as PCR.
Assuntos
Schistosoma japonicum/isolamento & purificação , Schistosoma/isolamento & purificação , Animais , Ensaio de Imunoadsorção Enzimática , Técnica Indireta de Fluorescência para Anticorpo , Humanos , Reação em Cadeia da PolimeraseRESUMO
Schistosomiasis is an acute and chronic parasitic disease caused by blood flukes (trematode worms) of the genus Schistosoma. Schistosoma japonicum (S. japonicum) infection has decreased significantly in prevalence and intensity of infection in China. However, this disease still remains a serious public health problem in some endemic areas of the Philippines and Indonesia. Thus, more accurate and sensitive methods are much needed for further control of this disease. Here, we review the research progress in techniques for the diagnosis of S. japonicum infection.
Assuntos
Esquistossomose Japônica/diagnóstico , Animais , Anticorpos Anti-Helmínticos/sangue , Antígenos de Helmintos , China/epidemiologia , DNA de Helmintos/análise , Humanos , Indonésia , Filipinas/epidemiologia , Reação em Cadeia da Polimerase , Prevalência , Schistosoma japonicum/genética , Schistosoma japonicum/imunologia , Schistosoma japonicum/isolamento & purificação , Esquistossomose Japônica/epidemiologia , Esquistossomose Japônica/parasitologia , Testes SorológicosRESUMO
Endothelin receptors (ETRs) are activated by vasoactive peptide endothelins and involved in the pathogenesis of hepatic fibrosis. However, less is known about the role of ETRs in Schistosoma (S.) japonicum-induced hepatic fibrosis. Here, we show that the expression of ETRs is markedly enhanced in the liver and spleen tissues of patients with schistosome-induced fibrosis, as well as in murine models. Additional analyses have indicated that the expression levels of ETRs in schistosomiasis patients are highly correlated with the portal vein and spleen thickness diameter, both of which represent the severity of fibrosis. Splenomegaly is a characteristic symptom of schistosome infection, and splenic abnormality may promote the progression of hepatic fibrosis. We further demonstrate that elevated levels of ETRs are predominantly expressed on splenic B cells in spleen tissues during infection. Importantly, using a well-studied model of human schistosomiasis, we demonstrate that endothelin receptor antagonists can partially reverse schistosome-induced hepatic fibrosis by suppressing the activation of splenic B cells characterized by interleukin-10 (IL-10) secretion and regulatory T (Treg) cell-inducing capacity. Our study provides insights into the mechanisms by which ETRs regulate schistosomiasis hepatic fibrosis and highlights the potential of endothelin receptor antagonist as a therapeutic intervention for fibrotic diseases.
Assuntos
Linfócitos B/patologia , Fibrose/patologia , Hepatopatias/patologia , Receptores de Endotelina/metabolismo , Schistosoma japonicum/isolamento & purificação , Esquistossomose/complicações , Baço/patologia , Adulto , Idoso , Animais , Linfócitos B/metabolismo , Linfócitos B/parasitologia , Feminino , Fibrose/etiologia , Fibrose/metabolismo , Humanos , Hepatopatias/etiologia , Hepatopatias/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Pessoa de Meia-Idade , Receptores de Endotelina/genética , Esquistossomose/parasitologia , Baço/metabolismo , Baço/parasitologiaRESUMO
BACKGROUND: Schistosomiasis japonica is an infectious disease caused by Schistosoma japonicum that seriously endangers human health. Domestic animals have important roles in disease transmission and goats are considered a primary reservoir host and source of infection. The prevalence and intensity of schistosomiasis infections have significantly decreased in China, and a more sensitive, specific detection method is urgently needed. The aim of this study was to develop a real-time PCR assay for accurate detection of S. japonicum infection in goats. METHODS: A real-time PCR method for detecting schistosomiasis japonica in goats was developed by amplification of a specific S. japonicum DNA fragment, and validated using a total of 94 negative and 159 positive plasma and serum samples collected in our previous study of S. japonicum infection. Both plasma and serum samples were evaluated by real-time PCR and enzyme-linked immunosorbent assay (ELISA). In addition, 120 goat plasma samples from an S. japonicum-endemic area (Wangjiang) and 33 from a non-endemic region (Weihai) were collected and evaluated using our method. RESULTS: The sensitivity and specificity of the real-time PCR for detecting infected samples were 98.74% (157/159, 95% CI: 95.53-99.85%) and 100% (94/94, 95% CI: 96.15-100%), respectively. For the ELISA, sensitivity and specificity were 98.11% (156/159, 95% CI: 94.59-99.61%) and 90.43% (85/94, 95% CI: 82.60-95.53%), respectively. Further, we found positivity rates for S. japonicum infection in Wangjiang and Weihai of 8.33% (10/120, 95% CI: 4.07-14.79%) and 0% (0/33, 95% CI: 0-10.58%), respectively. CONCLUSIONS: The results of this study indicate that our real-time PCR method exhibits higher sensitivity and specificity than ELISA and is a useful method for detection of S. japonicum infection in goats.
Assuntos
Cabras/parasitologia , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Schistosoma japonicum/isolamento & purificação , Esquistossomose Japônica/veterinária , Animais , China , DNA de Helmintos , Reservatórios de Doenças , Ensaio de Imunoadsorção Enzimática , Doenças das Cabras/diagnóstico , Humanos , Schistosoma japonicum/genética , Esquistossomose Japônica/diagnóstico , Sensibilidade e Especificidade , Zoonoses/diagnósticoRESUMO
Currently the diagnosis of schistosomiasis is mainly determined by observing the presence of eggs in host stool samples. Because of the overwhelming number of impurities in the stool, eggs are rarely observed. Therefore, the stool hatching method is used to observe the miracidia in the water. However, the miracidia of Schistosoma japonicum are small and difficult to detect, and missed detection is likely to occur when the infection level is low. In this study, recombinant streptococcal protein G-enhanced green fluorescent protein (rSPG-EGFP) was expressed, purified, and used as a fluorescence staining reagent for miracidia. A preliminary miracidium fluorescence staining method based on antigen and antibody bindingwas established by combining recombinant protein staining with the stool hatching method. The stool hatching method was used to collect the miracidia of S. japonicum, and Schistosoma-positive serum and the recombinant protein were mixed to assess the feasibility of fluorescence staining of miracidia. The miracidia of S. japonicum and Schistosoma turkestanicum were incubated with S. japonicum-positive serum and S. turkestanicum-positive serum, respectively, to identify miracidia species. When the fluorescence staining method was used to observe living miracidia, the miracidiawere labelled by the recombinant protein, and their motility status was not affected.
Assuntos
Schistosoma japonicum/isolamento & purificação , Esquistossomose Japônica/diagnóstico , Coloração e Rotulagem , Animais , Fluorescência , HumanosRESUMO
China once suffered greatly from schistosomiasis japonica, a major zoonotic disease. Nearly 70 years of multidisciplinary efforts have achieved great progress in disease control, with infections in both humans and bovines significantly reduced to very low levels. However, reaching for the target of complete interruption of transmission at the country level by 2030 still faces great challenges, with areas of ongoing endemicity and/or re-emergence within previously 'eliminated' regions. The objectives of this study were, by using meta-analytical methods, to estimate the overall prevalence of Schistosoma japonicum infections in abundant commensal rodent species in mainland China after the introduction of praziquantel for schistosomiasis treatment in humans and bovines in 1980s. In doing so we thereby aimed to further assess the role of wild rodents as potential reservoirs in ongoing schistosome transmission. Published studies on infection prevalence of S. japonicum in wild rodents in mainland China since 1980 were searched across five electronic bibliographic databases and lists of article references. Eligible studies were selected based on inclusion and exclusion criteria. Risks of within and across study biases, and the variations in prevalence estimates attributable to heterogeneities were assessed. The pooled infection prevalence and its 95% confidence intervals (CIs) were calculated with the Freeman-Tukey double arcsine transformation. We identified a total of 37 relevant articles involving 61 field studies which contained eligible data on 8,795 wild rodents across mainland China. The overall pooled infection prevalence was 3.86% (95% CI: 2.16-5.93%). No significant change in the overall pooled prevalence was observed between 1980-2003 (n = 23 studies) and 2004-current (n = 38 studies). However, whilst the estimated prevalence decreased over time in the marshland and lake regions, there was an apparent increase in prevalence within hilly and mountainous regions. Among seven provinces, a significant prevalence reduction was only seen in Jiangsu where most endemic settings are classified as the marshland and lakes. These estimates changed over season, ranging from 0.58% in spring to 22.39% in winter, in association with increases in rodent density. This study systematically analyzed S. japonicum infections in wild rodents from the published literature over the last forty years after the introduction of praziquantel for schistosomiasis treatment in humans and bovines in 1980s. Although numbers of schistosomiasis cases in humans and bovines have been greatly reduced, no such comparable overall change of infection prevalence in rodents was detected. Furthermore, there appeared to be an increase in S. japonicum prevalence in rodents over time within hilly and mountainous regions. Rodents have been projected to become the dominant wildlife in human-driven environments and the main reservoir of zoonotic diseases in general within tropical zones. Our findings thus suggest that it is now necessary to include monitoring and evaluation of potential schistosome infection within rodents, particularly in hilly and mountainous regions, if we are ever to reach the new 2030 elimination goals and to maximize the impact of future public, and indeed One Health, interventions across, regional, national and international scales.
Assuntos
Doenças dos Roedores/parasitologia , Esquistossomose Japônica/parasitologia , Esquistossomose Japônica/veterinária , Animais , Animais Selvagens/parasitologia , China/epidemiologia , Humanos , Praziquantel/administração & dosagem , Doenças dos Roedores/tratamento farmacológico , Doenças dos Roedores/epidemiologia , Roedores/parasitologia , Schistosoma japonicum/efeitos dos fármacos , Schistosoma japonicum/genética , Schistosoma japonicum/isolamento & purificação , Schistosoma japonicum/fisiologia , Esquistossomose Japônica/tratamento farmacológico , Esquistossomose Japônica/epidemiologiaRESUMO
Schistosomiasis occurs in 240 million people worldwide and is a major public health concern. Thus, early diagnosis and monitoring of schistosomiasis progression are needed to treat patients. Cell-free DNA (cfDNA) is present as fragments of parasite-derived DNA in host body fluids. Detection of this cfDNA in host blood may be a promising diagnostic marker of schistosomiasis. Therefore, in this study, we investigated the potential of internal transcribed spacer 2 (ITS2), a molecular taxonomy and barcoding marker, in diagnosing schistosomiasis using infected rabbit and mice sera. A 192 bp fragment of ITS2 was detected in the serum-isolated DNA from the infected host on different days after infection. We also determined the sensitivity of detecting ITS2 in mice with varying numbers of cercaria: cfDNA was present even in mice with low abundance of the parasite. Overall, our results show that cfDNA may be a potential tool for the early diagnosis and therapeutic evaluation of S. japonicum infection.
Assuntos
Ácidos Nucleicos Livres/sangue , Schistosoma japonicum/genética , Esquistossomose Japônica/genética , Esquistossomose Japônica/parasitologia , Esquistossomose/diagnóstico , Esquistossomose/genética , Soro/parasitologia , Animais , Biomarcadores/sangue , Cercárias/isolamento & purificação , DNA Intergênico/sangue , Humanos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Técnicas de Diagnóstico Molecular , Coelhos , Schistosoma japonicum/isolamento & purificação , CaramujosRESUMO
The Schistosomiasis Consortium for Operational Research and Evaluation (SCORE) was funded in 2008 to conduct research that would support country schistosomiasis control programs. As schistosomiasis prevalence decreases in many places and elimination is increasingly within reach, a sensitive and specific test to detect infection with Schistosoma mansoni and Schistosoma haematobium has become a pressing need. After obtaining broad input, SCORE supported Leiden University Medical Center (LUMC) to modify the serum-based antigen assay for use with urine, simplify the assay, and improve its sensitivity. The urine assay eventually contributed to several of the larger SCORE studies. For example, in Zanzibar, we demonstrated that urine filtration, the standard parasite egg detection diagnostic test for S. haematobium, greatly underestimated prevalence in low-prevalence settings. In Burundi and Rwanda, the circulating anodic antigen (CAA) assay provided critical information about the limitations of the stool-based Kato-Katz parasite egg-detection assay for S. mansoni in low-prevalence settings. Other SCORE-supported CAA work demonstrated that frozen, banked urine specimens yielded similar results to fresh ones; pooling of specimens may be a useful, cost-effective approach for surveillance in some settings; and the assay can be performed in local laboratories equipped with adequate centrifuge capacity. These improvements in the assay continue to be of use to researchers around the world. However, additional work will be needed if widespread dissemination of the CAA assay is to occur, for example, by building capacity in places besides LUMC and commercialization of the assay. Here, we review the evolution of the CAA assay format during the SCORE period with emphasis on urine-based applications.
Assuntos
Antígenos de Helmintos/imunologia , Glicoproteínas/imunologia , Proteínas de Helminto/imunologia , Schistosoma/imunologia , Esquistossomose/diagnóstico , Animais , Biomarcadores , Burundi/epidemiologia , Criança , Testes Diagnósticos de Rotina , Fezes/parasitologia , Feminino , Humanos , Testes Imunológicos , Masculino , Modelos Animais , Papio/parasitologia , Contagem de Ovos de Parasitas , Prevalência , Ruanda/epidemiologia , Santa Lúcia/epidemiologia , Schistosoma/isolamento & purificação , Schistosoma haematobium/imunologia , Schistosoma haematobium/isolamento & purificação , Schistosoma japonicum/imunologia , Schistosoma japonicum/isolamento & purificação , Schistosoma mansoni/imunologia , Schistosoma mansoni/isolamento & purificação , Esquistossomose/epidemiologia , Sensibilidade e Especificidade , Tanzânia/epidemiologia , Urina/parasitologiaRESUMO
BACKGROUND: Schistosomiasis is a neglected tropical parasitic disease associated with severe pathology, mortality and economic loss worldwide. Programs for disease control may benefit from specific and sensitive diagnostic methods to detect Schistosoma trematodes in aquatic environments. Here we report the development of novel environmental DNA (eDNA) qPCR assays for the presence of the human-infecting species Schistosoma mansoni, S. haematobium and S. japonicum. METHODOLOGY/PRINCIPAL FINDINGS: We first tested the specificity of the assays across the three species using genomic DNA preparations which showed successful amplification of target sequences with no cross amplification between the three focal species. In addition, we evaluated the specificity of the assays using synthetic DNA of multiple Schistosoma species, and demonstrated a high overall specificity; however, S. japonicum and S. haematobium assays showed cross-species amplification with very closely-related species. We next tested the effectiveness of the S. mansoni assay using eDNA samples from aquaria containing infected host gastropods, with the target species revealed as present in all infected aquaria. Finally, we evaluated the effectiveness of the S. mansoni and S. haematobium assays using eDNA samples from eight discrete natural freshwater sites in Tanzania, and demonstrated strong correspondence between infection status established using eDNA and conventional assays of parasite prevalence in host snails. CONCLUSIONS/SIGNIFICANCE: Collectively, our results suggest that eDNA monitoring is able to detect schistosomes in freshwater bodies, but refinement of the field sampling, storage and assay methods are likely to optimise its performance. We anticipate that environmental DNA-based approaches will help to inform epidemiological studies and contribute to efforts to control and eliminate schistosomiasis in endemic areas.
Assuntos
DNA Ambiental/isolamento & purificação , Água Doce/parasitologia , Schistosoma/classificação , Schistosoma/genética , Schistosoma/isolamento & purificação , Animais , DNA de Helmintos/isolamento & purificação , Monitoramento Ambiental , Genes de Helmintos/genética , Técnicas de Amplificação de Ácido Nucleico/veterinária , Filogenia , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Schistosoma haematobium/genética , Schistosoma haematobium/isolamento & purificação , Schistosoma japonicum/genética , Schistosoma japonicum/isolamento & purificação , Schistosoma mansoni/genética , Schistosoma mansoni/isolamento & purificação , Esquistossomose/epidemiologia , Esquistossomose/parasitologia , Esquistossomose mansoni/epidemiologia , Esquistossomose mansoni/parasitologia , Caramujos/parasitologia , Especificidade da Espécie , TanzâniaRESUMO
In recent years, the prevalence and infection intensity of Schistosoma japonicum in endemic areas of the Philippines have significantly decreased due to yearly population-based treatment strategies, yet transmission rates remain high and uninterrupted. An important indicator of active disease transmission is the presence of Schistosoma japonicum and its snail intermediate host Oncomelania hupensis quadrasi in freshwater habitats. In this study, we sought to apply a species-specific real-time PCR (qPCR) assay for the detection of S. japonicum and O. hupensis quadrasi in freshwater samples using environmental DNA approach that can complement the commonly utilized malacological survey in determining potential transmission foci in order to have a more effective snail surveillance strategy for schistosomiasis japonica in endemic areas. The newly developed assay was specific to S. japonicum and O. hupensis quadrasi with no amplification detected against non-target trematode Fasciola spp. and snails such as Lymnaea spp., Pomacea canaliculata, and Melanoides spp. that typically co-exist in the same environment. The assay effectiveness was determined using 19 environmental water samples collected from Northern Samar (N = 5 sites), Leyte (N = 11 sites) and Compostela Valley (N = 3 sites) and compared to malacological survey for determining O. hupensis quadrasi snail colonies and snail crushing to visualize S. japonicum cercariae. TaqMan qPCR targeting a short fragment of the cytochrome c oxidase subunit 1 (cox1) gene was positive for S. japonicum in 9 sites, for O. hupensis quadrasi in 9 sites, and for both S. japonicum and O. hupensis quadrasi in 5 sampling sites. Moreover, it was able to detect O. hupensis quadrasi in 3 out of 12 sites found negative and 6 out of 7 sites found positive through malacological survey, and in 4 of the 5 snail sites positive for snails with cercariae. Overall, this method can complement malacological surveys for monitoring of schistosomes in endemic areas of the Philippines, especially those with high risk of human infection.