Duplex reverse transcriptase PCR for concurrent detection of porcine astrovirus and porcine kobuvirus

Abstract

Pig farming plays a vital role in economic growth. Unfortunately, the swine farming sector of India has experienced a decline, partially due to several viral and bacterial infections. These diseases have direct impact on animal health, productivity, mortality rates, and economic returns. Viral gastroenteritis, caused by enteric viruses, is a severe condition that primarily results in infection in neonatal animals. Many enteric viruses are responsible for viral gastroenteritis amongst which Porcine astrovirus (PAstV) and Porcine kobuvirus (PKV) are the emerging ones. Current diagnostic approaches are serological tests, conventional PCR, and other molecular diagnostics, which are reliable but take time and are expensive for high throughput screening. To overcome these challenges a duplex RT-PCR assay was developed for identification of PAstV and PKV in pigs to boost up the intensity of diagnosis. This approach allowed rapid and inexpensive testing, which is particularly useful for the swine population. A duplex PCR assay showed sensitivity to a reliable range of concentrations for PAstV at an LOD of 2.74 ng and for PKV at 30 fg. This developed assay also found to be specific and did not detect other porcine viruses like PCV and PPV. The applicability of assay was also done for screening (n=50) pig samples collected from different regions of Punjab, India, and the developed nucleo-diagnostic identified both viruses (PAstV and PKV) at a single stance and thus enhanced control measures that would reduce losses occasioned by these diseases in the swine industry.