Development and Application of nanoPCR Method for Detection of Feline Panleukopenia Virus

SIMPLE SUMMARY: Feline panleukopenia is a severe infectious disease caused by the feline panleukopenia virus. It has caused great obstacles to the breeding of pet cats and the protection of rare feline animals such as lions and tigers. There is currently no effective therapy for FPV infection. The virus is extensively dispersed in the environment, and infected animals expel the virus in their feces, allowing the virus to spread; young kittens are especially vulnerable. Infection in kittens frequently is acute, and some cats may die unexpectedly without any signs, posing a significant challenge to early clinical identification. Some of the existing detection methods, such as qPCR, immunohistochemical analysis, colloidal gold test strips, etc., are time-consuming, laborious, costly, and difficult to operate, which is not conducive to popularization. Although colloidal gold test strips are fast and convenient, they have low sensitivity. A nanoPCR reaction was successfully created in this work by adding gold nanoparticles to the traditional PCR reaction system, and was capable of detecting FPV infection in the clinical situation. ABSTRACT: Feline panleukopenia (FP) is a severe viral illness caused by the feline panleukopenia virus (FPV), putting sectors like companion cat breeding and endangered feline conservation at risk. The virus has a high morbidity and fatality rate and is found all over the world. We created a novel FPV assay using nanoPCR technology and assessed the method’s specificity and sensitivity. The approach amplified a 345 bp nucleic acid fragment with a minimum detection limit of 7.97 × 10(2) copies/μL, which is about 100 times greater than traditional PCR. We collected anal swabs from 83 cats suspected of FPV infection for practical application, and the FPV-positive rate determined by the nanoPCR approach was 77.1%. In conclusion, the approach is more sensitive than conventional PCR and more convenient and cost-effective than qPCR methodology and may be utilized for the clinical detection of FPV.