Molecular characterization of rifampin-resistant isolates of Mycobacterium tuberculosis from Hungary by DNA sequencing and the line probe assay
Molecular characterization of rifampin-resistant isolates of Mycobacterium tuberculosis from Hungary by DNA sequencing and the line probe assay
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DOI：
10.1128/jcm.39.10.3736-3739.2001
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发表时间：
2001-10-01
影响因子：
9.4
通讯作者：
Salfinger, M
中科院分区：
文献类型：
--
作者：
Bártfai, Z;Somoskövi, A;Salfinger, M
对来自匈牙利3个县患者的29株耐利福平结核分枝杆菌（比例法测定）进行了与利福平耐药相关的两个rpoB区测序。29株耐药菌株中，27株在81-bp区（26株）或n端区（1株）发生突变，其余2株在这两个区域均未发生突变。突变的位置和频率与之前报道的不同。在这项研究中，最常见的突变是D516V，在38%的匈牙利菌株中发现，其频率比其他国家的研究中发现的频率高2到10倍。这29个分离株也用inin - lipa Rif进行了评估。TB试验（LiPA），一种快速检测利福平耐药性的反向杂交试验。虽然LiPA检测到26株耐药菌株存在rpoB突变，但4株菌株的突变类型无法确定，因为存在的突变不在LiPA试纸上包含的突变中。此外，一株利福平敏感对照菌株的沉默突变被LiPA解释为利福平耐药。这些发现表明，在将该检测纳入常规诊断工作之前，通过与每个地理位置的DNA序列结果进行比较来验证该快速分子检测的重要性。
Two regions of rpoB associated with rifampin resistance were sequenced in 29 rifampin-resistant (determined by the proportion method) isolates of Mycobacterium tuberculosis obtained from patients from three counties in Hungary. Of the 29 resistant strains, 27 had a mutation in either the 81-bp region (26 strains) or the N-terminal region (1 strain), while the other 2 strains had no mutations in either region. The locations and frequencies of the mutations differed from those previously reported. The most common mutation in this study, D516V, was found in 38% of the Hungarian strains, a frequency 2 to 10 times higher than that found in studies from other countries. These same 29 isolates were also evaluated with the Inno-LiPA Rif. TB test (LiPA), a reverse hybridization assay for the rapid detection of rifampin resistance. Although LiPA detected the presence of an rpoB mutation in 26 of the resistant isolates, the type of mutation could not be determined in 4 isolates because the mutations present were not among those included on the LiPA strip. In addition, a silent mutation in one of the rifampin-susceptible control strains was interpreted as rifampin resistant by LiPA. These findings demonstrate the importance of validating this rapid molecular test by comparison with DNA sequence results in each geographic location before incorporating the test into routine diagnostic work.