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Abstract
A diagnostic method for simultaneously detecting and distinguishing African Swine Fever (ASF), porcine circovirus type 2 (PCV2), and porcine parvovirus (PPV) in clinical specimens is critical for differential diagnosis, monitoring, and control in the field. Three primer pairs were designed and used to create a multiplex PCR assay. In addition, 356 porcine post mortem tissue samples from various parts of India's North Eastern region were tested by the developed multiplex PCR assay to demonstrate its accuracy. Using the designed primers, each of the ASF, PCV2 and PPV target genes was amplified, but no other porcine virus genes were detected. The assay's limit of detection was 102 copies/µl of PCV2, PPV, or ASFV. The detection of PCV2, PPV, and ASF in postmortem tissue samples revealed that they are co-circulating in India's North-Eastern region. The percentage positivity (PP) for PCV2, PPV and ASF single infection were 7.02% (25/356), 3.93% (14/356), and 3.37% (12/356), respectively, while the PP for PCV2& PPV co-infection was 2.80% (10/356), ASF & PCV2 co infection was 1.4% (5/356) and the ASF, PPV& PCV2 co-infection was1.40% (5/356). The results also indicate that the ASF can infect pigs alongside PCV and PPV.
Disclosure statement
No potential conflict of interest was reported by the author(s).
Author contributions
SRP, RD, PJD and GSS conceived of and designed the experiments, and revised the manuscript. AKY and SR Validation of the assay. VKG final approval of the manuscript. All authors reviewed and approved the manuscript.
Acknowledgement
The authors are thankful to the Director, ICAR-National Research Center on Pig, Guwahati, Assam, India for providing necessary facilities to carry out the present research. The authors are also thankful to the Department of Biotechnology, Government of India for the financial aid for the present study under the project entitled “Swinostics” (BT/PR41886/NER/95/1720/2021).