J Vet Sci.  2006 Jun;7(2):181-187. 10.4142/jvs.2006.7.2.181.

Random amplified polymorphic DNA (RAPD) analysis of Mycobacterium tuberculosis strains in India

Affiliations
  • 1Division of Bacteriology and Mycology, Indian Veterinary Research Institute, Izatnagar-243122, U.P., India. rishendra_verma@yahoo.com
  • 2Division Veterinary Biotechnology, Indian Veterinary Research Institute, Izatnagar-243122, U.P., India.

Abstract

The usefulness of random amplification of polymorphic DNA (RAPD) analysis for typing Indian strains of M. tuberculosis was investigated. M. tuberculosis H37Rv, M. tuberculosis DT and 42 clinical isolates of M. tuberculosis were subjected to RAPD-PCR using 7 random decamer primers. All 7 primers were found to be differentiated and produced specific RAPD profiles. The polymorphic amplicons served as RAPD markers for M. tuberculosis. The dendrograms, obtained by different primers, showed the discriminatory ability of the primers. RAPD analysis provided a rapid and easy means of identifying polymorphism in M. tuberculosis isolates, and it was found to be a valuable alternative epidemiological tool. In addition, the
results
of the present study showed heterogeneity in the M. tuberculosis strains in the population studied.

Keyword

Mycobacterium tuberculosis; RAPD; typing

MeSH Terms

Animals
Cattle
Cattle Diseases/microbiology
DNA, Bacterial/*analysis/genetics
Gene Expression Profiling
Humans
India/epidemiology
Lung/microbiology
Mycobacterium tuberculosis/*genetics/*isolation&purification
Phylogeny
*Random Amplified Polymorphic DNA Technique
Sputum/microbiology
Swine
Swine Diseases/microbiology
Tuberculosis, Pulmonary/*epidemiology/*microbiology/veterinary
Variation (Genetics)

Figure

  • Fig. 1 RAPD profiles of Mycobacterium tuberculosis strains with primer OPN-02. (A) Lane M-100 bp DNA marker ladder; 1-(162/ 93); 2-(186/96); 3-(12/87); 4-(321/96); 5-(380/98); 6-(191/94); 7-(199/94); 8-(197/94); 9-(193/94); 10-(425/2); 11-H37Rv; 12-DT; Lane M2-50 bp DNA marker ladder. (B) Lane M-100 bp DNA marker ladder; 1-(11/S); 2-(3/S); 3-(6/S); 4-(5/S); 5-(9/S); 6-(SpS10); 7-(SpS11); 8-(SpS16); 9-(SpS22); 10-(SpS8); 11-(SpS19); 12-(SpS17); 13-(SpS14); 14-(SpS13); 15-(SpS4); 16-(SpS6); 17- H37Rv; 18- DT; Lane M2-50 bp DNA marker ladder. (C) Lane M-100 bp DNA marker ladder; 1-(45/90); 2-(2/S); 3-(4/S); 4-(8/S); 5-(7/S); 6-(10/ S); 7-(12/S); 8-(13/S); 9-(14/S); 10-(15/S); 11-(1/86); 12-(203/94); 13-(125/92); 14-(320/96); 15-(439/1); 16-(128/92); 17-H37Rv; 18-DT; Lane M2-50 bp DNA marker ladder.

  • Fig. 2 Dendrogram showing genetic relatedness among Mycobacterium tuberculosis strains with primer OPN-02.


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