A continuation: study and characterisation of Mycobacterium leprae short tandem repeat genotypes and transmission of leprosy in Cebu, Philippines.
A continuation: study and characterisation of Mycobacterium leprae short tandem repeat genotypes and transmission of leprosy in Cebu, Philippines.
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发表时间：
2009-09
期刊：
影响因子：
0.6
通讯作者：
R. Sakamuri;Jordan D. Harrison;R. Gelber;P. Saunderson;P. Brennan;M. Balagon;V. Vissa
中科院分区：
文献类型：
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作者：
R. Sakamuri;Jordan D. Harrison;R. Gelber;P. Saunderson;P. Brennan;M. Balagon;V. Vissa
目的研究菲律宾宿务麻风患者麻风分枝杆菌串联重复位点的稳定性和等位基因多样性，探讨多位点可变串联重复位点（VNTR）分析（MLVA）分型检测传播的适用性。方法在宿务Leonard Wood Memorial皮肤诊所就诊的70例新诊断麻风患者，从患者的裂隙皮肤涂片（SSS）刮片中提取总DNA，采用单位点或多重PCR扩增13个麻风分枝杆菌VNTR位点。通过DNA测序或片段长度分析方法获得每个VNTR位点的重复数。分子流行病学数据库中包括了医疗、社会和地理细节。结果与结论与单短串联重复序列（STR）扩增和DNA测序相比，多重PCR （MP）和片段长度分析（FLA）方法更有效、准确。来自四个不同样本的患者内部MLVA模式在微卫星中被保守，而在5名患者中的4名患者中观察到一个或多个多态和口吃易发微卫星的差异。这13个基因座可以区分宿务麻风分枝杆菌菌株，但在多病例家庭的潜伏期和个体间传播过程中，MLVA模式足够稳定。因此，麻风分枝杆菌MLVA在宿雾地区具有菌株分型和传播研究的潜力。
OBJECTIVE To study the stability and allelic diversity of tandem repeat loci in M. leprae in leprosy patients of Cebu, Philippines, and the suitability of multilocus variable number of tandem repeat (VNTR) analysis (MLVA) typing for detecting transmission. METHODS Seventy newly diagnosed leprosy patients consulting at the Leonard Wood Memorial, Cebu Skin Clinic Total DNA was extracted from slit skin smear (SSS) scrapings of each patient and used for amplification of 13 M. leprae VNTR loci by single locus or multiplex PCR. Number of repeats for each VNTR locus was obtained by DNA sequencing or fragment length analysis methods. Medical, social and geographic details were included in the molecular epidemiology database. RESULTS AND CONCLUSIONS Multiplex PCR (MP) and fragment length analysis (FLA) methods were found to be more efficient and accurate compared to single short tandem repeat (STR) amplification and DNA sequencing. Intra-patient MLVA patterns from four different samples were conserved in the minisatellites, while differences in one or more of the polymorphic and stutter prone microsatellites was observed, in four of five patients. The 13 loci could differentiate M. leprae strains in Cebu, however, MLVA patterns were stable enough during incubation and transmission between individuals within multi-case families. Thus M. leprae MLVA has potential for strain typing and transmission studies in Cebu.