{"title":"一种新型的半巢式PCR检测方法，具有高灵敏度和强大的特异性，可以同时检测人和动物的bocavavirus。","authors":"Chutchai Piewbang, Aisyah Nikmatuz Zahro, Panida Poonsin, Jiratchaya Puenpa, Cherdpong Phupolphan, Nathamon Kosoltanapiwat, Thongchai Ngamprasertwong, Julien Claude, Porntippa Lekchareonsuk, Yong Poovorawan, Somporn Techangamsuwan","doi":"10.1093/infdis/jiaf090","DOIUrl":null,"url":null,"abstract":"<p><strong>Background: </strong>Bocaviruses (BoVs), belonging to the Parvoviridae family, pose significant challenges in detection due to their genetic diversity and cross-species transmission capabilities. Efficient and broad-spectrum detection methods are essential for understanding BoV epidemiology and addressing potential zoonotic risks.</p><p><strong>Methods: </strong>We developed a seminested polymerase chain reaction (PCR) assay for simultaneous detection of diverse BoV species across human and animal hosts. Primers were designed by analyzing 765 BoV genome sequences, targeting conserved regions spanning the NP1 to VP2 genes. Sensitivity was determined through analytical tests, and specificity was evaluated against 39 non-BoV viruses. Validation was performed using spiked biological samples, and the method was applied to 552 clinical samples from 542 hosts, encompassing a broad range of mammalian species.</p><p><strong>Results: </strong>The assay demonstrated high sensitivity, detecting BoVs at concentrations as low as 0.2 copies/µL. Specificity tests confirmed no cross-reactivity with other viral families. Validation using 37 strains representing 29 BoV species affirmed its broad efficacy. BoVs were identified across diverse hosts, including humans, bats, canines, porcines, rodents, and felines. Additionally, novel host associations were observed, such as Panthera uncia bocaparvovirus (PuBoV) in a tiger and serval cat, canine bocavirus 2 (CBoV-2) in raccoon dogs, and feline bocaviruses (FBoV) in murid rodents. Human bocaviruses were also detected in monkey samples, indicating potential pathogen spillover.</p><p><strong>Conclusions: </strong>This seminested PCR method provides a sensitive and specific tool for BoV detection, enhancing surveillance in human and animal populations. It is instrumental in monitoring zoonotic risks and emerging infectious threats, offering critical insights into BoV epidemiology and cross-species transmission dynamics.</p>","PeriodicalId":50179,"journal":{"name":"Journal of Infectious Diseases","volume":" ","pages":"499-509"},"PeriodicalIF":4.5000,"publicationDate":"2025-08-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"A Novel Seminested Polymerase Chain Reaction Assay With High Sensitivity and Robust Specificity Enables Simultaneous Detection of Human and Animal Bocaviruses.\",\"authors\":\"Chutchai Piewbang, Aisyah Nikmatuz Zahro, Panida Poonsin, Jiratchaya Puenpa, Cherdpong Phupolphan, Nathamon Kosoltanapiwat, Thongchai Ngamprasertwong, Julien Claude, Porntippa Lekchareonsuk, Yong Poovorawan, Somporn Techangamsuwan\",\"doi\":\"10.1093/infdis/jiaf090\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Background: </strong>Bocaviruses (BoVs), belonging to the Parvoviridae family, pose significant challenges in detection due to their genetic diversity and cross-species transmission capabilities. Efficient and broad-spectrum detection methods are essential for understanding BoV epidemiology and addressing potential zoonotic risks.</p><p><strong>Methods: </strong>We developed a seminested polymerase chain reaction (PCR) assay for simultaneous detection of diverse BoV species across human and animal hosts. Primers were designed by analyzing 765 BoV genome sequences, targeting conserved regions spanning the NP1 to VP2 genes. Sensitivity was determined through analytical tests, and specificity was evaluated against 39 non-BoV viruses. Validation was performed using spiked biological samples, and the method was applied to 552 clinical samples from 542 hosts, encompassing a broad range of mammalian species.</p><p><strong>Results: </strong>The assay demonstrated high sensitivity, detecting BoVs at concentrations as low as 0.2 copies/µL. Specificity tests confirmed no cross-reactivity with other viral families. Validation using 37 strains representing 29 BoV species affirmed its broad efficacy. BoVs were identified across diverse hosts, including humans, bats, canines, porcines, rodents, and felines. Additionally, novel host associations were observed, such as Panthera uncia bocaparvovirus (PuBoV) in a tiger and serval cat, canine bocavirus 2 (CBoV-2) in raccoon dogs, and feline bocaviruses (FBoV) in murid rodents. Human bocaviruses were also detected in monkey samples, indicating potential pathogen spillover.</p><p><strong>Conclusions: </strong>This seminested PCR method provides a sensitive and specific tool for BoV detection, enhancing surveillance in human and animal populations. It is instrumental in monitoring zoonotic risks and emerging infectious threats, offering critical insights into BoV epidemiology and cross-species transmission dynamics.</p>\",\"PeriodicalId\":50179,\"journal\":{\"name\":\"Journal of Infectious Diseases\",\"volume\":\" \",\"pages\":\"499-509\"},\"PeriodicalIF\":4.5000,\"publicationDate\":\"2025-08-14\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of Infectious Diseases\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://doi.org/10.1093/infdis/jiaf090\",\"RegionNum\":2,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"IMMUNOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Infectious Diseases","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1093/infdis/jiaf090","RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"IMMUNOLOGY","Score":null,"Total":0}
A Novel Seminested Polymerase Chain Reaction Assay With High Sensitivity and Robust Specificity Enables Simultaneous Detection of Human and Animal Bocaviruses.
Background: Bocaviruses (BoVs), belonging to the Parvoviridae family, pose significant challenges in detection due to their genetic diversity and cross-species transmission capabilities. Efficient and broad-spectrum detection methods are essential for understanding BoV epidemiology and addressing potential zoonotic risks.
Methods: We developed a seminested polymerase chain reaction (PCR) assay for simultaneous detection of diverse BoV species across human and animal hosts. Primers were designed by analyzing 765 BoV genome sequences, targeting conserved regions spanning the NP1 to VP2 genes. Sensitivity was determined through analytical tests, and specificity was evaluated against 39 non-BoV viruses. Validation was performed using spiked biological samples, and the method was applied to 552 clinical samples from 542 hosts, encompassing a broad range of mammalian species.
Results: The assay demonstrated high sensitivity, detecting BoVs at concentrations as low as 0.2 copies/µL. Specificity tests confirmed no cross-reactivity with other viral families. Validation using 37 strains representing 29 BoV species affirmed its broad efficacy. BoVs were identified across diverse hosts, including humans, bats, canines, porcines, rodents, and felines. Additionally, novel host associations were observed, such as Panthera uncia bocaparvovirus (PuBoV) in a tiger and serval cat, canine bocavirus 2 (CBoV-2) in raccoon dogs, and feline bocaviruses (FBoV) in murid rodents. Human bocaviruses were also detected in monkey samples, indicating potential pathogen spillover.
Conclusions: This seminested PCR method provides a sensitive and specific tool for BoV detection, enhancing surveillance in human and animal populations. It is instrumental in monitoring zoonotic risks and emerging infectious threats, offering critical insights into BoV epidemiology and cross-species transmission dynamics.
期刊介绍：
Published continuously since 1904, The Journal of Infectious Diseases (JID) is the premier global journal for original research on infectious diseases. The editors welcome Major Articles and Brief Reports describing research results on microbiology, immunology, epidemiology, and related disciplines, on the pathogenesis, diagnosis, and treatment of infectious diseases; on the microbes that cause them; and on disorders of host immune responses. JID is an official publication of the Infectious Diseases Society of America.
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