With a population of more than 231 million, Indonesia carries the fifth highest TB （tuberculosis） burden globally. Therefore, many efforts to support TB control program are being undertaken. One of them is strengthe...With a population of more than 231 million, Indonesia carries the fifth highest TB （tuberculosis） burden globally. Therefore, many efforts to support TB control program are being undertaken. One of them is strengthening the capabilities in molecular analysis of the resistance of TB bacteria to the drugs such as ETH （ethionamide）. ETH is prodrug that needs to be activated by mycobacterial enzymes in order to exert their antimicrobial activity. The aim of this study was to understand the molecular basis for Mycobacterium tuberculosis resistance to ETH. DNA （Deoxyribonucleic acid） extraction of M. tuberculosis was amplified with PCR （polymerase chain reaction） on ethA gene and analyzed their drug resistance by using SSCP （single strand conformation polymorphism） technique. The DNA of M. tuberculosis （H37Rv） strain was used as positive control. Of 53 extracts of DNA amplified, 11 （20.75%） were positive for the target gene and none of samples was resistent to ETH based on the alteration of DNA band mobility shift on acrylamide gel. This low number of positive PCR amplicons may be related to annealing temperature and initial DNA content in specimen. Study on the implementation of hot-SSCP by labeling the DNA with 32p radioisotope shows that this nuclear based technique has the higher quality in term of the visualization.展开更多
文摘With a population of more than 231 million, Indonesia carries the fifth highest TB （tuberculosis） burden globally. Therefore, many efforts to support TB control program are being undertaken. One of them is strengthening the capabilities in molecular analysis of the resistance of TB bacteria to the drugs such as ETH （ethionamide）. ETH is prodrug that needs to be activated by mycobacterial enzymes in order to exert their antimicrobial activity. The aim of this study was to understand the molecular basis for Mycobacterium tuberculosis resistance to ETH. DNA （Deoxyribonucleic acid） extraction of M. tuberculosis was amplified with PCR （polymerase chain reaction） on ethA gene and analyzed their drug resistance by using SSCP （single strand conformation polymorphism） technique. The DNA of M. tuberculosis （H37Rv） strain was used as positive control. Of 53 extracts of DNA amplified, 11 （20.75%） were positive for the target gene and none of samples was resistent to ETH based on the alteration of DNA band mobility shift on acrylamide gel. This low number of positive PCR amplicons may be related to annealing temperature and initial DNA content in specimen. Study on the implementation of hot-SSCP by labeling the DNA with 32p radioisotope shows that this nuclear based technique has the higher quality in term of the visualization.
