Mokrousov Igor, Filliol Ingrid, Legrand Eric, Sola Christophe, Otten Tatiana, Vyshnevskaya Elena, Limeschenko Elena, Vyshnevskiy Boris, Narvskaya Olga, Rastogi Nalin
Unité de la Tuberculose et des Mycobactéries, Institut Pasteur de Guadeloupe, Morne Jolivière, Pointe-à-Pitre.
Res Microbiol. 2002 May;153(4):213-9. doi: 10.1016/s0923-2508(02)01311-6.
This investigation evaluated the potential of RNA/RNA mismatch analysis for the detection of rifampin resistance among 38 multiple-drug-resistant (MDR) isolates of Mycobacterium tuberculosis from northwestern Russia. The results obtained were compared with a commercialized line probe assay and rpoB sequencing, and the genetic diversity of the isolates was also investigated in parallel using spoligotyping and variable number of tandem DNA repeats (VNTR). The mismatch analysis revealed 3 distinct RNA cleavage profiles permitting the subdivision of the strains into mutation groups 1 to 3, the most common being group 1 (28 of 38 isolates) that contained a majority of strains with a TCG531>TTG (Ser->Leu) mutation, followed by group 2 (6 of 38 isolates) characterized by different mutations in the codon CAC526 (His), and group 3 (4 of 38 isolates), all characterized by a GAC516(Asp) mutation. Spoligotyping revealed the Beijing type to be the most prevalent among mismatch group 1 (24 out of 28 strains), suggesting that the most frequent rpoB mutation among the Beijing family in our setting was TCG531 >TTG (Ser->Leu). All the Beijing type isolates were also characterized by a unique VNTR pattern made up of exact tandem repeats (ETR)-A to E of 42435. We conclude that the Beijing genotype constitutes the major family of MDR-TB isolates currently circulating in northwestern Russia, and that the in-house RNA/RNA mismatch analysis may be successfully used for rapid and reliable diagnosis of rifampin-resistant tuberculosis in this setting.
本研究评估了RNA/RNA错配分析在检测俄罗斯西北部38株多重耐药结核分枝杆菌分离株中利福平耐药性的潜力。将所得结果与商业化的线性探针分析和rpoB测序进行比较，并同时使用间隔寡核苷酸分型（spoligotyping）和可变数目串联重复序列（VNTR）对分离株的遗传多样性进行研究。错配分析揭示了3种不同的RNA切割图谱，可将菌株分为突变组1至3，最常见的是组1（38株分离株中的28株），其中大多数菌株具有TCG531>TTG（Ser->Leu）突变，其次是组2（38株分离株中的6株），其特征是密码子CAC526（His）发生不同突变，以及组3（38株分离株中的4株），均以GAC516（Asp）突变为特征。间隔寡核苷酸分型显示北京型在错配组1中最为普遍（28株菌株中的24株），这表明在我们的研究环境中，北京家族中最常见的rpoB突变是TCG531>TTG（Ser->Leu）。所有北京型分离株还具有由42435的精确串联重复序列（ETR）-A至E组成的独特VNTR模式。我们得出结论，北京基因型是目前在俄罗斯西北部流行的多重耐药结核分枝杆菌分离株的主要家族，并且内部RNA/RNA错配分析可成功用于在这种情况下快速可靠地诊断利福平耐药结核病。
