Establishment and application of triple PCR method for FAstV, FPV and FECoV

ABSTRACT

Feline Astrovirus (FAstV), Feline Parvovirus(FPV) and Feline Enteric Coronavirus (FECoV) are important pathogens causing diarrhea in cats.In order to establish a molecular detection method which can differentiate the three pathogens in the same PCR system, an FAstV/FPV/FECoV triple PCR method was established with optimized primer concentrations and annealing temperature, and specificity, sensitivity and repeatability were tested. The results showed that the PCR method could only identify FAstV (320 bp), FPV (468 bp) and FECoV (664 bp) genes, while not other canine and feline related pathogens. The detection limits of FAstV, FPV and FECoV were 2x10~7 copy/L (7.1 pg/L),4.7x10~6 copy/L (2.4 pg/L) and 7x10~6 copy/L (5.1 pg/L) respectively. The established triple PCR method was used to detect 207 cat fecal samples collected in Chengdu from 2019 to 2020, including 141 diarrhea samples and 66 clinical health samples. The detection rates of FAstV, FPV and FECoV were 24.15% (50/207), 37.20% (77/207) and 15.46% (32/207) respectively, and the co-infection rates of FAstV/FPV, FPV/FECoV and FAstv/FECoV were 9.18%,6.28% and 6.28% respectively. In conclusion, the triple PCR method of FAstV/FPV/FECoV was successfully established, and could be applied for virus detection and epidemiological investigation.