VNTR typing of Mycobacterium leprae in South Indian leprosy patients.
VNTR typing of Mycobacterium leprae in South Indian leprosy patients.
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DOI：
10.47276/lr.80.3.290
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发表时间：
2009-09
期刊：
影响因子：
0.6
通讯作者：
V. Shinde;H. Newton;R. Sakamuri;V. Reddy;Suman Jain;Abraham Joseph;T. Gillis;I. Nath;G. Norman;V. Vissa
中科院分区：
文献类型：
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作者：
V. Shinde;H. Newton;R. Sakamuri;V. Reddy;Suman Jain;Abraham Joseph;T. Gillis;I. Nath;G. Norman;V. Vissa
目的研究短串联重复序列(STR)基因组标记对印度南部安得拉邦和泰米尔纳德邦麻风患者麻风分枝杆菌菌株变异的适用性、稳定性和多样性。材料与方法从新诊断的麻风患者皮损和不同部位采集狭缝皮肤涂片(SSS)。除了5名患者外，每个患者的SSSS都被汇集在一起。从SSS样本中提取总DNA。用多重聚合酶链式反应(MP)或单一聚合酶链式反应方法从DNA中扩增出麻风杆菌STR。通过片段长度分析(FLA)或DNA测序获得每个STR基因座(STR等位基因)的重复数。结果与结论多重聚合酶链式反应最大限度地减少了DNA和试剂的使用，与FLA一起用于STR菌株分型是一种既省时又省钱的方法。通过对13个STR基因座的南印度分离株的检测，确定(AC)8b、(GGT)5、6-3a(RpoT)、21-3、27-5和23-3等位基因在两个研究群体中是保守的。在来自安得拉邦的一个家庭中，两名患者的麻风杆菌STR模式在18个表明共同感染源的基因座中有16个是相同的。在泰米尔纳德邦测试的5名患者中，15个STR基因座中有14个没有患者内部变异。总之，这些研究表明STR毒株分型适用于评估短程传播链。
OBJECTIVES To study the suitability, stability and diversity of short tandem repeat (STR) genomic markers to elicit strain variation in the Mycobacterium leprae isolates within leprosy patients from Andhra Pradesh and Tamil Nadu states in South India. MATERIALS AND METHODS Slit skin smear (SSS) samples were collected from lesions and various body sites of newly diagnosed leprosy patients. The SSSs from each patient were pooled, except in the case of five patients. Total DNA was extracted from SSS samples. M. leprae STRs were amplified from the DNA either by multiplex PCR (MP) or single PCR methods. The number of repeats for each STR locus (the STR allele) was obtained either by fragment length analysis (FLA) or by DNA sequencing of the PCR amplicons. RESULTS AND CONCLUSION Multiplex PCR minimised the use of DNA and reagents, and together with FLA, was time and cost effective for STR strain typing. After examination of the isolates of South Indian origin at 13 STR loci, it was determined that the alleles for (AC)8b, (GGT)5, 6-3a (rpoT), 21-3, 27-5, and 23-3 were conserved in two study populations. In a family from Andhra Pradesh, the M. leprae STR patterns in two patients were identical in 16 of 18 loci which indicate a common source of infection. Fourteen of 15 STR loci showed no intra-patient variation in the five patients tested in Tamil Nadu. Altogether, these studies indicate the suitability of STR strain typing for assessing short-range transmission chains.