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Abstract
Mycobacteria are aerobic, nonspore forming, non-motile,single-cell bacteria.Of more than 40 currently recognized species of mycobacteria, Mycobacterium tuberculosis, the causative agent of human TB is the commonest pathogen for pulmonary and extra pulmonary tuberculosis cases. The other members of the Mycobacterium tuberculosis complex (MTC) or the nontubercular mycobacterium (NTM) produces similar diseases which cannot be differentiated from tuberculosis by clinical symptoms and signs. But this differentiation is important as the chemotherapy varies widely according to the strain of mycobacterium. The burden of morbidity and mortality of tuberculosis is rapidly growing worldwide, particularly with the HIV/AIDS epidemic. The strain identification of Mycobacterium remains a cumbersome, labor intensive and expensive procedure, which requires 3 to 12 weeks of time. The conventional methods of strain identification lack proper standardization and precise diagnosis. The prime objective of this study is to overcome these problems.A multiplex PCR using 3 amplicons of 165,365, and 541 base pair target sequences was done with a total number of 165 clinical isolates of suspected Koch’s patients. Strain identification was compared both by conventional methods and multiplex PCR. The results of the study show that this multiplex PCR is supposed to be less complicated, less time consuming, cost-effective and superior to the conventional methods. It is also applicable for culture negative samples where strain identification is not possible by conventional approach.
Highlights
Based on surveillance and survey data, WHO estimates that 9.27 million new cases of TB occurred in 2007.Of these 9.27 million new cases, an estimated 44% were new smear positive cases, 14.8% were HIV positive cases and 4.9% were MDR-TB
An estimated 1.32 million HIVnegative people died from TB in 2007 and there were an additional 4,56,000 TB deaths among HIV positive people [ICD-10]
Incidence of MDR-TB in INDIA is reported to vary from 0.1%-5.4%.MDR-TB among all new TB cases are 2.8%, whereas 17% among the previously treated cases [1,2,3,4,5]
Summary
Based on surveillance and survey data, WHO estimates that 9.27 million new cases of TB occurred in 2007.Of these 9.27 million new cases, an estimated 44% were new smear positive cases, 14.8% were HIV positive cases and 4.9% were MDR-TB (which includes 3.1% new cases and 19% previously treated cases). India is ranking top among 27 high MDR-TB burden countries. Incidence of MDR-TB in INDIA is reported to vary from 0.1%-5.4%.MDR-TB among all new TB cases are 2.8%, whereas 17% among the previously treated cases [1,2,3,4,5]. Among the mycobacterial diseases (both pulmonary and extrapulmonary) M.tuberculosis complex has got the lion’s share. The infection with Nontubercular Mycobacterium (NTM) or Mycobacterium Other Than Tuberculosis (MOTT) is increasing rapidly
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