Spoligotyping for Identification of Pulmonary Tuberculosis with Type 2 Diabetes Mellitus Profiles in Warangal, T.S.

Background: The prevalence of pulmonary tuberculosis infections had increased among the type 2 Diabetes Mellitus (DM) patients in India, although the biological basis underlying this susceptibility remains poorly characterized. Based on the symptom’s, chest X-rays and their correlation with clinical and diagnostic parameters, we had suspected diabetic-tuberculosis co-infections. As Chest XRay plays a vital role in the identification of pulmonary tuberculosis infections, we emphasized more on it. Chest X-ray features included were; (infiltrate, cavitation, miliary shadows, pleural effusion, mediastinal lymphadenopathy and extent of lesions) such features were analyzed to identify associations with biological/clinical parameters through univariate and multivariate logistic regression. Method: The study was carried out in diabetic type 2 patients suspected with pulmonary tuberculosis infections, Warangal, India. Spoligotyping was used for identification, detection and characterization of M. tuberculosis complex bacteria in clinical samples. This method is based on polymerase chain reaction (PCR) amplification of a highly polymorphic direct repeat locus in the M. tuberculosis genome. It is helpful in detecting causative bacteria and in providing epidemiologic information on genotyping strain identities. Results: Based on the chest X-Ray of 200 diabetic patients suspected with pulmonary infections, we found 26 infiltrates, 30 cavitation, 28 miliary shadows, 35 pleural effusion, 46 mediastinal lymphadenopathy and 35 were confirmed for the extent of lesions, which supported us to further screen for pulmonary tuberculosis of 200 subjects tested, 113 were males, 85 were females and 2 were children. All 200 subjects were tested for pulmonary tuberculosis, 36 were positive by smear microscopy and 20 were culture positive. Phenotypic and genotypic variations were found for all the 20 identified clinical isolates, by conventional and molecular methods of 20 clinical isolates, 4 MDR-TB were identified based on the Drug Susceptibility Test for first-line drugs. Of 20 clinical isolates, we took 10 clinical isolates (4-were MDR-TB and 6- were MTB) and 1- was control sample of H37RV used for spoligotyping and showed different patterns Bejing (1) and Lineages of East Asian, of family EA13(2) and Lineages of Indo-Oceanic, of family LAM1(1) and Lineages of Euro-American and 6 were found to be MTB of family 33(2) and Lineages of Indo-Oceanic, of family CAS(4) and Lineages of Un-known family. CAS (Central Asian) of M.tuberculosis strains showed more prevailing spoligotype pattern in Diabetic Pulmonary Tuberculosis patients. Conclusion: Implementing such a method in clinical settings would be useful in surveillance of tuberculosis transmission and in interventions to prevent further spread of this disease among the Diabetic Pulmonary Tuberculosis co-infections.