The Rapid Identification of Atypical Mycobacterium in Pulmonary Tuberculosis (PTB) Patients: Evaluation of QUB۳۲۳۲ Locus Using the VNTR Method
- سال انتشار: 1388
- محل انتشار: مجله علمی پژوهشی دانشگاه علوم پزشکی زنجان، دوره: 17، شماره: 67
- کد COI اختصاصی: JR_ZUMS-17-67_004
- زبان مقاله: انگلیسی
- تعداد مشاهده: 291
نویسندگان
Azad University of Tehran, Tehran, Iran
Mycobacteriology Research Center (MRC), National Research Institute of Tuberculosis and Lung Disease (NRITLD), Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Dept. of Microbiology, Azad University of Tehran, Iran
Dept. of Biology, Azad University of Tehran, Iran
Dept. of Microbiology, Azad University of Jahrom, Jahrom, Iran
National Research Institute of Tuberculosis and Lung Disease (NRITLD), Shahid Beheshti University of Medical Sciences, Tehran, Iran
National Research Institute of Tuberculosis and Lung Disease (NRITLD), Shahid Beheshti University of Medical Sciences, Tehran, Iran
چکیده
Background and Objective: Identification of atypical mycobacterium (Non tuberculosis Mycobacterium NTM) is important because of the worldwide propagation of these organisms. Recently, molecular studies have identified the specific loci for mycobacterium species by DNA - finger printing methods, but these methods are time-consuming and expensive. In this study, in addition to hsp۶۵ PCR-RFLP method, QUB۳۲۳۲ locus was evaluated for differentiation of atypical mycobacterium from mycobacterium tuberculosis complex. Materials and Methods: This study was performed on ۳۷۱ pulmonary and non pulmonary specimens separated from patients with the symptoms of pulmonary tuberculosis (PTB). After the isolation and culturing of mycobacterium strains using the Lowenstein Jensen media, biochemical tests including production of Niacin, Catalase activity, Nitrate reduction, pigment production and growth rate were performed. Drug susceptibility testing was performed by proportional method. DNA extraction was performed by phenol-chloroform method. hsp۶۵ gene was amplified by PCR. Subsequently the amplicons were digested with three restriction enzymes namely AvaII, HphI and HpaII and electrophoresed on ۳% agarose gel. QUB۳۲۳۲ locus was also evaluated for differentiation of atypical mycobacterium and mycobacterium tuberculosis complex. Results: Out of ۳۷۱ isolates, ۳۲ (۸.۶%) were multi-drug resistant TB (MDR-TB), ۱۸۴ (۴۹.۵%) were susceptible and ۱۵۵ (۴۲.۵%) were non MDR (combined resistance) that ۱۵% of MDR cases and ۲۵% of non MDR cases were non tuberculosis mycobacterium. Out of ۳۱ slow growing isolates, ۵۸% were M. simiae and ۱۹% were M. kansasii. The sensitivity of QUB۳۲۳۲ locus for differentiation of the atypical mycobacterium from mycobacterium tuberculosis complex was ۸۰%. From the total of ۴۳ NTM samples, ۱۲ (۲۷.۹%) were rapid growing and ۷۲% were slow growing. Conclusion: QUB۳۲۳۲ locus has the high discriminative power for differentiation of atypical mycobacterium from the mycobacterium tuberculosis complex, therefore, it can be used as a substitute for PCR-RFLP method.کلیدواژه ها
Key words: Atypical mycobacterium, VNTR, QUB۳۲۳۲ locusاطلاعات بیشتر در مورد COI
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