Development of a Taqman-based real-time PCR assay for detection of porcine sapelovirus infection in pigs. Issue 3 (3rd May 2020)

• Record Type: Journal Article
• Authors: Kumari, Swati; Ray, P. K.; Singh, Rahul; Desingu, P. A.; Sharma, G. Taru; Saikumar, G.
• Is Part Of: Animal biotechnology. Volume 31:Issue 3(2020)
• Page Start: 264
• Page End: 267
• Publication Date: 2020-05-03
• Subjects: PSV -- Taqman probe -- plasmid -- cloned; Animal biotechnology -- Periodicals; Animals, Domestic -- genetics -- Periodicals; Animals, Genetically Modified -- Periodicals; 660.6
• DOI: 10.1080/10495398.2018.1549561
• Languages: English
• ISSNs: 1049-5398

Abstract

Abstract: The objective of the present study was to develop a rapid, simple, specific and sensitive Taqman-based real-time PCR assay for porcine sapelovirus (PSV) detection. Specific primers and probe were designed from the five untranslated regions (UTRs) of the viral genome. The detection limit of the real-time PCR was 10 2 copies. The specificity of the Taqman real-time PCR assay was evaluated using other animal viruses and nuclease free water as a negative control. Strong fluorescent signals were obtained only in the detection of PSV real-time PCR and conventional RT-PCR were preformed simultaneously on 90 faecal samples. Based on conventional RT-PCR study 17.7% (16/90) of the faecal samples were positive for PSV. Whereas 21 of 90 samples (23.3%) were positive by real-time RT-PCR. The results showed that real-time PCR was more sensitive than the conventional RT-PCR assay. In conclusion, the Taqman real-time PCR assay for detection of PSV developed, herein, is sensitive, specific, and reliable. This assay will be useful for clinical diagnosis, epidemiological, and pathogenesis studies.