The aim of this study was to develop a novel duplex nested RT-PCR (dnRT-PCR) method which can simultaneously detect and distinguish feline herpesvirus type 1 (FHV-1) and feline calicivirus (FCV) in cats with upper respiratory tract disease (URTD) and evaluate the performance of this assay in clinical screening. Four pairs of primers based on TK gene of FHV-1 and ORF2 gene of FCV were designed, respectively, and a dnRT-PCR assay was developed and evaluated. A total of 115 clinical swaps of cats collected from private veterinary clinics and animal shelters in Jilin province between November 2016 and May 2017 were used to evaluate this newly developed dnRT-PCR method compared with conventional assay. The results indicated that the detection limits of this method were 10^1.0 TCID₅₀/ml, 10^2.2 TCID₅₀/ml and 10^2.0 TCID₅₀/ml for FHV-1, FCV and a mixture of the two viruses which were 100-fold and 1000-fold higher than single and duplex conventional RT-PCR/PCR, respectively, and was also highly specific. Clinical screening indicated the concordance rates for dnRT-PCR and single conventional RT-PCR/PCR were 99.13% (kappa=0.955) for FHV-1 and 98.26% (kappa=0.956) for FCV, respectively. These results suggest that the duplex nested RT-PCR is a rapid and sensitive method that is useful for clinical diagnosis and etiological investigation of feline caolicivirus and feline herpesvirus.

To Cite This Article: Yi S, Niu J, Dong H, Hu G, Guo Y, Wang K, Zhang S and Zhao Y, 2018. Rapid detection of feline calicivirus and feline herpesvirus by duplex nested RT-PCR. Pak Vet J, 38(4): 347-352. http://dx.doi.org/10.29261/pakvetj/2018.086