Development of Nested Polymerase Chain Reaction for Diagnosis of FeLV Infection in Cats

Abstract

The Nested Polymerase Chain Reaction test has been developed for a diagnosis of FeLV infection, by detecting proviral DNA of FeLV. In transient and latent infection cats, the freely circulating viral proteins present at high concentration in blood circulation within a few weeks to months. This latent infection is difficult to be detected by ELISA or ELISA application technique (Commercial test kit). The problem of latent infection status can be resolved by detection of proviral sequence which integrates in host genome. Nested PCR reaction was designed in two steps. First round, PCR produced 770 bp product and second round produced 601 bp product. The nested PCR method is more specific and sensitive than regular PCR method. Target gene of oligonucleotide primers is gag which could express very low rate of protein mutation. Therefore, the high mutation rates of retrovirus do not affect the specificity of this detection method.