Development of a quadruple PCR-based gene microarray for detection of vaccine and wild-type classical swine fever virus, African swine fever virus and atypical porcine pestivirus
Abstract Background Classical swine fever (CSF), African swine fever (ASF), and atypical porcine pestivirus (APPV) are acute, virulent, and contagious viral diseases currently hampering the pig industry in China, which result in mummification or stillbirths in piglets and mortality in pigs. Diagnost...
| Main Authors: | , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2022-11-01
|
| Series: | Virology Journal |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12985-022-01933-9 |
| _version_ | 1828174092650414080 |
|---|---|
| author | Ying-ju Xia Lu Xu Jun-jie Zhao Yuan-xi Li Rui-zhi Wu Xiang-peng Song Qi-zu Zhao Ye-bing Liu Qin Wang Qian-yi Zhang |
| author_facet | Ying-ju Xia Lu Xu Jun-jie Zhao Yuan-xi Li Rui-zhi Wu Xiang-peng Song Qi-zu Zhao Ye-bing Liu Qin Wang Qian-yi Zhang |
| author_sort | Ying-ju Xia |
| collection | DOAJ |
| description | Abstract Background Classical swine fever (CSF), African swine fever (ASF), and atypical porcine pestivirus (APPV) are acute, virulent, and contagious viral diseases currently hampering the pig industry in China, which result in mummification or stillbirths in piglets and mortality in pigs. Diagnostic assays for the differentiation of infection and vaccination of CSFV, in addition to the detection of ASFV and APPV, are urgently required for better prevention, control, and elimination of these viral diseases in China. Methods A quadruple PCR-based gene microarray assay was developed in this study to simultaneously detect wild-type and vaccine CSFV strains, ASFV and APPV according to their conserved regions. Forty-two laboratory-confirmed samples, including positive samples of 10 other swine viral diseases, were tested using this assay to confirm its high specificity. Results This assay's limit of detections (LODs) for the wild-type and vaccine CSFV were 6.98 and 6.92 copies/µL. LODs for ASFV and APPV were 2.56 × 10 and 1.80 × 10 copies/µL, respectively. When compared with standard RT-PCR or qPCR for CSFV (GB/T 26875–2018), ASFV (MARR issue No.172), or APPV (CN108611442A) using 219 clinical samples, the coincidence was 100%. The results showed that this assay with high sensitivity could specifically distinguish ASFV, APPV, and CSFV, including CSFV infection and immunization. Conclusion This assay provides a practical, simple, economic, and reliable test for the rapid detection and accurate diagnosis of the three viruses and may have good prospects for application in an epidemiological investigation, prevention, and control and elimination of these three diseases. |
| first_indexed | 2024-04-12T04:07:26Z |
| format | Article |
| id | doaj.art-6eeb01156cb347d39a386212a0fc3162 |
| institution | Directory Open Access Journal |
| issn | 1743-422X |
| language | English |
| last_indexed | 2024-04-12T04:07:26Z |
| publishDate | 2022-11-01 |
| publisher | BMC |
| record_format | Article |
| series | Virology Journal |
| spelling | doaj.art-6eeb01156cb347d39a386212a0fc31622022-12-22T03:48:35ZengBMCVirology Journal1743-422X2022-11-0119111110.1186/s12985-022-01933-9Development of a quadruple PCR-based gene microarray for detection of vaccine and wild-type classical swine fever virus, African swine fever virus and atypical porcine pestivirusYing-ju Xia0Lu Xu1Jun-jie Zhao2Yuan-xi Li3Rui-zhi Wu4Xiang-peng Song5Qi-zu Zhao6Ye-bing Liu7Qin Wang8Qian-yi Zhang9China Institute of Veterinary Drug ControlChina Institute of Veterinary Drug ControlChina Institute of Veterinary Drug ControlChina Institute of Veterinary Drug ControlChina Institute of Veterinary Drug ControlChina Institute of Veterinary Drug ControlChina Institute of Veterinary Drug ControlChina Institute of Veterinary Drug ControlChina Institute of Veterinary Drug ControlChina Institute of Veterinary Drug ControlAbstract Background Classical swine fever (CSF), African swine fever (ASF), and atypical porcine pestivirus (APPV) are acute, virulent, and contagious viral diseases currently hampering the pig industry in China, which result in mummification or stillbirths in piglets and mortality in pigs. Diagnostic assays for the differentiation of infection and vaccination of CSFV, in addition to the detection of ASFV and APPV, are urgently required for better prevention, control, and elimination of these viral diseases in China. Methods A quadruple PCR-based gene microarray assay was developed in this study to simultaneously detect wild-type and vaccine CSFV strains, ASFV and APPV according to their conserved regions. Forty-two laboratory-confirmed samples, including positive samples of 10 other swine viral diseases, were tested using this assay to confirm its high specificity. Results This assay's limit of detections (LODs) for the wild-type and vaccine CSFV were 6.98 and 6.92 copies/µL. LODs for ASFV and APPV were 2.56 × 10 and 1.80 × 10 copies/µL, respectively. When compared with standard RT-PCR or qPCR for CSFV (GB/T 26875–2018), ASFV (MARR issue No.172), or APPV (CN108611442A) using 219 clinical samples, the coincidence was 100%. The results showed that this assay with high sensitivity could specifically distinguish ASFV, APPV, and CSFV, including CSFV infection and immunization. Conclusion This assay provides a practical, simple, economic, and reliable test for the rapid detection and accurate diagnosis of the three viruses and may have good prospects for application in an epidemiological investigation, prevention, and control and elimination of these three diseases.https://doi.org/10.1186/s12985-022-01933-9Classical swine feverAfrican swine feverAtypical pestivirusPolymerase chain reactionDifferential diagnosisGene chip |
| spellingShingle | Ying-ju Xia Lu Xu Jun-jie Zhao Yuan-xi Li Rui-zhi Wu Xiang-peng Song Qi-zu Zhao Ye-bing Liu Qin Wang Qian-yi Zhang Development of a quadruple PCR-based gene microarray for detection of vaccine and wild-type classical swine fever virus, African swine fever virus and atypical porcine pestivirus Virology Journal Classical swine fever African swine fever Atypical pestivirus Polymerase chain reaction Differential diagnosis Gene chip |
| title | Development of a quadruple PCR-based gene microarray for detection of vaccine and wild-type classical swine fever virus, African swine fever virus and atypical porcine pestivirus |
| title_full | Development of a quadruple PCR-based gene microarray for detection of vaccine and wild-type classical swine fever virus, African swine fever virus and atypical porcine pestivirus |
| title_fullStr | Development of a quadruple PCR-based gene microarray for detection of vaccine and wild-type classical swine fever virus, African swine fever virus and atypical porcine pestivirus |
| title_full_unstemmed | Development of a quadruple PCR-based gene microarray for detection of vaccine and wild-type classical swine fever virus, African swine fever virus and atypical porcine pestivirus |
| title_short | Development of a quadruple PCR-based gene microarray for detection of vaccine and wild-type classical swine fever virus, African swine fever virus and atypical porcine pestivirus |
| title_sort | development of a quadruple pcr based gene microarray for detection of vaccine and wild type classical swine fever virus african swine fever virus and atypical porcine pestivirus |
| topic | Classical swine fever African swine fever Atypical pestivirus Polymerase chain reaction Differential diagnosis Gene chip |
| url | https://doi.org/10.1186/s12985-022-01933-9 |
| work_keys_str_mv | AT yingjuxia developmentofaquadruplepcrbasedgenemicroarrayfordetectionofvaccineandwildtypeclassicalswinefevervirusafricanswinefevervirusandatypicalporcinepestivirus AT luxu developmentofaquadruplepcrbasedgenemicroarrayfordetectionofvaccineandwildtypeclassicalswinefevervirusafricanswinefevervirusandatypicalporcinepestivirus AT junjiezhao developmentofaquadruplepcrbasedgenemicroarrayfordetectionofvaccineandwildtypeclassicalswinefevervirusafricanswinefevervirusandatypicalporcinepestivirus AT yuanxili developmentofaquadruplepcrbasedgenemicroarrayfordetectionofvaccineandwildtypeclassicalswinefevervirusafricanswinefevervirusandatypicalporcinepestivirus AT ruizhiwu developmentofaquadruplepcrbasedgenemicroarrayfordetectionofvaccineandwildtypeclassicalswinefevervirusafricanswinefevervirusandatypicalporcinepestivirus AT xiangpengsong developmentofaquadruplepcrbasedgenemicroarrayfordetectionofvaccineandwildtypeclassicalswinefevervirusafricanswinefevervirusandatypicalporcinepestivirus AT qizuzhao developmentofaquadruplepcrbasedgenemicroarrayfordetectionofvaccineandwildtypeclassicalswinefevervirusafricanswinefevervirusandatypicalporcinepestivirus AT yebingliu developmentofaquadruplepcrbasedgenemicroarrayfordetectionofvaccineandwildtypeclassicalswinefevervirusafricanswinefevervirusandatypicalporcinepestivirus AT qinwang developmentofaquadruplepcrbasedgenemicroarrayfordetectionofvaccineandwildtypeclassicalswinefevervirusafricanswinefevervirusandatypicalporcinepestivirus AT qianyizhang developmentofaquadruplepcrbasedgenemicroarrayfordetectionofvaccineandwildtypeclassicalswinefevervirusafricanswinefevervirusandatypicalporcinepestivirus |