STANDARDISATION OF REAL-TIME PCR FOR THE DETECTION OF ACUTE CANINE LEPTOSPIROSIS

Abstract

Leptospirosis is one of the most widespread zoonoses. The early diagnosis of the disease based on the symptoms is often difficult because of its overlap ping symptoms with many other febrile illnesses. The present study envisages the evaluation of real time PCR and conventional PCR as compared to the Microscopic Agglutination Test (MAT), for the detection of acute canine leptospirosis. A total of 75 samples were collected from the cases suspected for canine leptospirosis, which include sera for conducting MAT, whole blood for PCR and Real time PCR and urine samples. The sensitivity, specificity and accuracy of conventional and real time PCR assays were determined keeping MAT as the reference. When the sera samples at 1:400 dilution were tested in MAT, 36 of them were found to be positive. The sensitivity, specificity and accuracy of the conventional PCR when compared to MAT was 80.55, 97.4 and 89 percent, respectively. Similarly the values observed for real time PCR vs MAT were 91.6, 84.61 and 88 percent, respectively. From the results, it could be concluded that real time PCR assay standardized in the present study was found to be more sensitive than the conventional PCR. In addition, the real time PCR assay could deliver the results quickly and enable quantification of the organisms in the sample, which would aid in much more efficient case management.

Keywords

• leptospirosis
• real-time pcr
• amplification plot
• melt curve