Detection of Leptospira in Canine Samples using Polymerase Chain Reaction based on LipL32gene and Biochemical Profiles in Canine Leptospirosis

Abstract

Canines face the risk of getting infected with commonly communicable diseases like leptospirosis, brucellosis, rabies, anthrax etc., Leptospirosis, a zoonoanthroponosis of worldwide distribution, is an acute febrile illness. Leptospirosis can be immune suppressive, predominantly human infection, commonly transmitted to human through contact of animal urine. We report here a case study of canines infected with leptospirosis, Leptospira has been isolated from blood. Leptospirosis is a septicemia bacterial disease affecting animals and human being, the infection may be symptomatic, mild or severe and acute or chronic. The clinical signals are often related to kidney disease, liver disease and other reproductive dysfunction. The disease currently diagnosed by Microscopic agglutination test (MAT). But, MAT is unable to differentiate the acute from chronic infection for the proper treatment. Since the disease affecting the kidney and the liver it is expected to have alterations in the biochemical profiles. AST and ALT in canine serum samples suspected for jaundice due to leptospirosis. In this study elevated AST and ALT were observed in positive patients. Hence, estimation of biochemical profiles may be used as an adjunct to MAT in diagnosis of acute infection. The present study was aimed to the polymerase chain reaction with the biochemical profiling for the diagnosis of leptospirosis. Biochemical and molecular techniques were performed on 40 samples of serum taken from dogs. Leptospira positives identified by dark field microscopy and PCR assay targeting partial LipL32gene of Leptospira using P23/24 primers, with a product size of 532 base pair long which are Leptospira specific. The amplicon subjected to restriction enzyme digestion using AluI, HinfI and ClaI for product of P23/24 and biochemical profile was made to correlate the biochemical AST and ALT with different clinical manifestation of leptospirosis in canines. Evidence of renal dysfunction in the present study is in conformity with the other studies. A combined effort of clinical expertise like molecular techniques along with the confirmatory laboratory back up. PCR was a reliable and precise diagnosis of leptospirosis when compare to biochemical technique in our study. The research presented here will be helpful to improve diagnosis and control of leptospirosis in other endemic region.