Daria L Manalo, Jude Karlo G Bolivar, Karl Ian T Ermino, Jeromir G Bondoc, Mark Joseph M Espino, Efraim P Panganiban, Kathyleen S Nogrado, Raffy Jay C Fornillos, Mario A Jiz, Lydia R Leonardo, Ian Kendrich C Fontanilla
{"title":"基于SYBR绿色的实时荧光定量PCR检测环境水样中湖北钉螺DNA的建立。","authors":"Daria L Manalo, Jude Karlo G Bolivar, Karl Ian T Ermino, Jeromir G Bondoc, Mark Joseph M Espino, Efraim P Panganiban, Kathyleen S Nogrado, Raffy Jay C Fornillos, Mario A Jiz, Lydia R Leonardo, Ian Kendrich C Fontanilla","doi":"10.3390/tropicalmed10050140","DOIUrl":null,"url":null,"abstract":"<p><p><i>Oncomelania hupensis quadrasi</i> is the intermediate host of <i>S. japonicum</i>, the causative species of schistosomiasis in the Philippines. Conventionally, risk areas are identified by procedures requiring highly skilled personnel and constant surveillance efforts. Recent developments in disease diagnostics explore the utilization of environmental DNA as targets for polymerase chain reactions in disease surveillance. In this study, a low-cost, specific, and efficient SYBR Green-based real-time PCR assay to detect <i>O. h. quadrasi</i> DNA from water samples was developed, optimized, and validated. Primers were designed based on the A18 microsatellite region of <i>O. h. quadrasi.</i> The assay exhibited a detection limit of one copy number per microliter at 99.4% efficiency and R<sup>2</sup> = 0.999, which specifically amplified <i>O. h. quadrasi</i> DNA only. Validation of this assay in environmental water samples demonstrated 100% PPV and NPV values, suggesting its potential as a tool for identifying risk areas, pathogen-directed surveillance, policy making, and disease control.</p>","PeriodicalId":23330,"journal":{"name":"Tropical Medicine and Infectious Disease","volume":"10 5","pages":""},"PeriodicalIF":2.8000,"publicationDate":"2025-05-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12115604/pdf/","citationCount":"0","resultStr":"{\"title\":\"Development of a SYBR Green-Based Real-Time PCR Assay to Detect <i>Oncomelania hupensis quadrasi</i> DNA in Environmental Water Samples.\",\"authors\":\"Daria L Manalo, Jude Karlo G Bolivar, Karl Ian T Ermino, Jeromir G Bondoc, Mark Joseph M Espino, Efraim P Panganiban, Kathyleen S Nogrado, Raffy Jay C Fornillos, Mario A Jiz, Lydia R Leonardo, Ian Kendrich C Fontanilla\",\"doi\":\"10.3390/tropicalmed10050140\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p><i>Oncomelania hupensis quadrasi</i> is the intermediate host of <i>S. japonicum</i>, the causative species of schistosomiasis in the Philippines. Conventionally, risk areas are identified by procedures requiring highly skilled personnel and constant surveillance efforts. Recent developments in disease diagnostics explore the utilization of environmental DNA as targets for polymerase chain reactions in disease surveillance. In this study, a low-cost, specific, and efficient SYBR Green-based real-time PCR assay to detect <i>O. h. quadrasi</i> DNA from water samples was developed, optimized, and validated. Primers were designed based on the A18 microsatellite region of <i>O. h. quadrasi.</i> The assay exhibited a detection limit of one copy number per microliter at 99.4% efficiency and R<sup>2</sup> = 0.999, which specifically amplified <i>O. h. quadrasi</i> DNA only. Validation of this assay in environmental water samples demonstrated 100% PPV and NPV values, suggesting its potential as a tool for identifying risk areas, pathogen-directed surveillance, policy making, and disease control.</p>\",\"PeriodicalId\":23330,\"journal\":{\"name\":\"Tropical Medicine and Infectious Disease\",\"volume\":\"10 5\",\"pages\":\"\"},\"PeriodicalIF\":2.8000,\"publicationDate\":\"2025-05-20\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12115604/pdf/\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Tropical Medicine and Infectious Disease\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://doi.org/10.3390/tropicalmed10050140\",\"RegionNum\":4,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"INFECTIOUS DISEASES\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Tropical Medicine and Infectious Disease","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.3390/tropicalmed10050140","RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"INFECTIOUS DISEASES","Score":null,"Total":0}
Development of a SYBR Green-Based Real-Time PCR Assay to Detect Oncomelania hupensis quadrasi DNA in Environmental Water Samples.
Oncomelania hupensis quadrasi is the intermediate host of S. japonicum, the causative species of schistosomiasis in the Philippines. Conventionally, risk areas are identified by procedures requiring highly skilled personnel and constant surveillance efforts. Recent developments in disease diagnostics explore the utilization of environmental DNA as targets for polymerase chain reactions in disease surveillance. In this study, a low-cost, specific, and efficient SYBR Green-based real-time PCR assay to detect O. h. quadrasi DNA from water samples was developed, optimized, and validated. Primers were designed based on the A18 microsatellite region of O. h. quadrasi. The assay exhibited a detection limit of one copy number per microliter at 99.4% efficiency and R2 = 0.999, which specifically amplified O. h. quadrasi DNA only. Validation of this assay in environmental water samples demonstrated 100% PPV and NPV values, suggesting its potential as a tool for identifying risk areas, pathogen-directed surveillance, policy making, and disease control.
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