Detecting parvovirus in dogs using PCR and cell cultures

Abstract

BACKGROUND: Thetype 2 is an acute viral disease in puppies and is considered a worldwide disease. AIM: This study was performed to identify the(CPV) throughout 2022 and 2023 in Iraq. The novelty of this study was to isolate this virus in two types of cell cultures (diploid and fibroblast cell cultures) and confirm the isolated virus via PCR technique. METHODS: Sixty fecal samples were taken from suspected infected dogs in the Baghdad Governorates who were clinically sick and thought to have CPV. The first performed qualitative antigen detection on the samples with a rapid test, then the positive samples were propagated on the feline kidney (FK) and embryonated chicken fibroblast cells and finally identified the isolated virus on six randomly selected samples by amplifying the VP2 gene with a conventional polymerase chain reaction. RESULTS: Rapid tests found 50% of fecal swabs as positive results. Additionally, the results of the cytopathic effect (CPE) at the second and third passages on chicken embryo and FK cell cultures revealed rounder and more detached cells. Furthermore, the isolated virus was confirmed through amplified the Vp2 gene of the virus by polymerase chain reaction. CONCLUSION: The focus of this study was to isolate of parvovirus that causes diarrhea in puppies, which is quite frequent in Iraq. The virus was isolated using both primary and secondary cell cultures. The solitary virus causes cells to become more differentiated and rounded due to CPEs. The virus was confirmed to be real using a polymerase chain reaction experiment. This study also suggests doing more epidemiological research and sequencing analysis to determine the prevalence of CPV serotypes in Iraq and the efficacy of trade in and locally made vaccinations in preventing against viral infection.