Year: 2017 | Month: August | Volume 7 | Issue 4
Molecular Detection of Mycobacterium bovis Targeting esxB (CFP-10) in Blood Samples and Lymph Node Aspirates by Conventional PCR and qRT-PCR TaqMan Assay
Abstract:
Bovine tuberculosis, a chronic infectious disease is caused by an intracellular acid-fast bacilli Mycobacterium bovis that affects broad range of mammalian hosts. CFP-10 is a 10 kDa secreted antigen coded by esxB gene located on RD1 region of genome and is responsible for virulence of Mycobacterium bovis. DNA extraction of blood (n=48) and lymph node aspirates (n=48) was done and extracted DNA was subjected to PCR by targeting esxB gene with band size of 302 bp. None of the blood sample and lymph node aspirates was positive for M. bovis with esxB gene by PCR. The sensitivity of esxB was 8 pg/μl by conventional PCR. Among 48 blood samples targeted for esxB (CFP-10) gene using In house developed primer probe mix, one sample (2.1%) whose CT was 34 was considered positive by real-time PCR. Out of 48 animals (lymph node aspirates), four samples (8.3%) whose CT was between 29-34 were considered positive by real-time PCR. Remaining samples whose CT values were equal to or greater than 35 were considered negative. The sensitivity of esxB was 800 fg/μl by real time PCR. This study indicates the diagnostic potential of esxB by using real time PCR TaqMan Assay
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