A Novel Seminested Polymerase Chain Reaction Assay With High Sensitivity and Robust Specificity Enables Simultaneous Detection of Human and Animal Bocaviruses
Issued Date
2025-08-15
Resource Type
ISSN
00221899
eISSN
15376613
Scopus ID
2-s2.0-105013258062
Pubmed ID
39969571
Journal Title
Journal of Infectious Diseases
Volume
232
Issue
2
Start Page
499
End Page
509
Rights Holder(s)
SCOPUS
Bibliographic Citation
Journal of Infectious Diseases Vol.232 No.2 (2025) , 499-509
Suggested Citation
Piewbang C., Zahro A.N., Poonsin P., Puenpa J., Phupolphan C., Kosoltanapiwat N., Ngamprasertwong T., Claude J., Lekchareonsuk P., Poovorawan Y., Techangamsuwan S. A Novel Seminested Polymerase Chain Reaction Assay With High Sensitivity and Robust Specificity Enables Simultaneous Detection of Human and Animal Bocaviruses. Journal of Infectious Diseases Vol.232 No.2 (2025) , 499-509. 509. doi:10.1093/infdis/jiaf090 Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/111754
Title
A Novel Seminested Polymerase Chain Reaction Assay With High Sensitivity and Robust Specificity Enables Simultaneous Detection of Human and Animal Bocaviruses
Corresponding Author(s)
Other Contributor(s)
Abstract
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.