University of Khartoum

Detection of Multi Drug Resistant TB Strains Using Conventional And Molecular Techniques

Detection of Multi Drug Resistant TB Strains Using Conventional And Molecular Techniques

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Title: Detection of Multi Drug Resistant TB Strains Using Conventional And Molecular Techniques
Author: Ismat, Gusai Hussein Abdel Samad
Abstract: Background: - Multidrug-resistance against the first line drugs for treatment of tuberculosis arises when the patients fail to complete the prescribed doses, and when the concentration of the drugs inside their bodies is not enough to kill all of the causative bacteria .This may be attributed to stoppage of treatment by patients when they feel better or they are unable to pay for the whole regimen that takes months. The infectivity of MDRT-TB strains is not affected and they remain to be fully virulent as non - resistant strains. The study aimed to detect the multi- drug resistant TB strains using conventional and molecular techniques and to detect the mutations that may be found in an effort to decrease the time of diagnosis and to have more accurate results that lead to fast and effective treatment. Methods:- The study was done on patients with pulmonary tuberculosis at Abu-Anga Teaching Hospital , Omdurman province during the year 2005. Samples of sputum from in-patients suffering from pulmonary tuberculosis were collected. These samples were pretreated with 4%NaOH and cultivated on L.J. media. Conventional methods (68Co labile catalase test , Nitrates reduction test , growth on PNB medium were used to identify the isolates . Collection , transport, pretreatment of samples , inoculation, incubation, reading and reporting of growth were done according to the WHO instructions, (1998). Susceptibility to rifampicin and isoniazid (proportion method) was performed for the previously obtained isolates.DNA from 20 resistant isolates was extracted . Molecular techniques such as PCR, cloning and sequencing were performed to detect the KatG gene responsible for isoniazid resistance. Results :- During the year 2005 , 6736 patients visited the out-patient clinic of Abu-Anga Hospital, 4699 were males and 2037 females. Only 431 patients (322 males, 109 females) were admitted to the hospital. Three hundred and five isolates were obtained, identified as Mycobacterium complex. The sensitivity test for the 305 isolates showed that 238(92.8%) isolates were sensitive to both rifampicin and isoniazid. Nineteen (6.2%) isolates were resistant to both rifampicin and isoniazid .Two isolates were resistant to isoniazid only and one isolate was resistant to rifampicin only. Multidrug resistance was found in 6.2%.of the isolates. Mutation of type S315T was observed in 20 isolates. Conclusion:- The prevalence of MDR-TB was found to be 6.2 % among clinical isolates at Abu-anga Hospital. The results were obtained in short time when using molecular techniques rather than conventional ones which take time . This provides quick results and fast treatment for MDR-TB patients.
URI: http://khartoumspace.uofk.edu/123456789/26722


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