Sensitivity Assay of Polymerase Chain Reaction for Detection of Canine Adeno Virus Infection in Dogs

Abstract

This study was done with the approach of finding out minimum detection limit of canine adeno virus in clinical samples using Polymerase chain reaction.A polymerase chain reaction was performed using reported primers for detection of Canine Adeno Virus (CAV) in the blood sample obtained from different regions of India. Detection and surveillance of desease very important for diagnosis and prevention of Disease.40 samples were screened for CAV infection in a time period of six months. DNA extracted from samples used as templet in PCR reaction using primers specific to E3 gene forward primer ICHA and reverse primer ICHB of CAV1 and CAV2 virus gives a band size of 508 bp which should be the size of amplified product. Positive sample of CAV in PCR were taken for study. Assay was performed using multidilution of extracted DNA of positive sample each dilution were used as template for PCR reaction .End point of the dilution obtained where no band was visible in agarose gel when visualize in geldoc system, same DNA was used for dilutions to check repeatability of PCR .Ten replicate PCR reactions were Studied. The detection limit find was at the dilution of 1:1000 is 0 .20 ng per ?l The study was valuable in determining the efficiency of PCR for detection of CAV Virus in clinical samples.

Key words:Canine Adeno Virus (CAV), reported primers, clinical samples, Infectious canine hepatitis, utilizing reported primers