Jadhav R S, Kamble R R, Shinde V S, Edward S, Edward V K
Stanley Browne Laboratory, Richardson Leprosy Hospital Sangli-Miraj Road, Miraj 416 410, Maharashtra, India.
Indian J Lepr. 2005 Apr-Jun;77(2):116-27.
The relevance of bacterial index (BI) for understanding the prognosis of leprosy patients on treatment has been extensively debated, as it does not give a very clear idea of the viability of the bacteria in patients under treatment. Here we used slit-skin smear samples of leprosy patients to test the suitability for studying viability of Mycobacterium leprae using reverse transcription polymerase chain reaction (RT-PCR). For this purpose, we recruited 13 multibacillary (MB) leprosy patients (8 lepromatous and 5 borderline lepromatous). Of these, 7 were relapse cases, 3 were under treatment (MB-MDT), 2 were new cases and 1 had completed treatment. We carried out extraction of RNA using Trizol reagent (Life Technologies, UK) from the slit-skin smear samples from these patients. The RNA preparation was then used for the RT-PCR using Mycobacterium leprae-specific primers for the fragment of 16s ribosomal RNA gene. Samples from both the new cases, 4 suspected relapse cases and 1 patient under treatment showed positive RT-PCR results. Other 6 patients whose smear samples did not show any amplification by RT-PCR were on MB-MDT from 8 to 30 months. The usefulness of the technique needs to be validated using mouse footpad technique and also should be more extensively explored for studying the viability of M. leprae, the efficacy of treatment and the presence of other mycobacteria in the slit-skin smear samples.
细菌指数（BI）对于理解正在接受治疗的麻风病患者的预后相关性一直存在广泛争议，因为它并不能非常清晰地反映接受治疗患者体内细菌的存活情况。在此，我们使用麻风病患者的皮肤涂片样本，通过逆转录聚合酶链反应（RT-PCR）来检测其用于研究麻风分枝杆菌存活情况的适用性。为此，我们招募了13例多菌型（MB）麻风病患者（8例瘤型和5例界线类偏瘤型）。其中，7例为复发病例，3例正在接受治疗（多菌型麻风联合化疗），2例为新发病例，1例已完成治疗。我们使用Trizol试剂（英国赛默飞世尔科技公司）从这些患者的皮肤涂片样本中提取RNA。然后将RNA制剂用于RT-PCR，使用针对16s核糖体RNA基因片段的麻风分枝杆菌特异性引物。新发病例、4例疑似复发病例和1例正在接受治疗的患者的样本RT-PCR结果均为阳性。其他6例皮肤涂片样本经RT-PCR未显示任何扩增的患者正在接受8至30个月的多菌型麻风联合化疗。该技术的实用性需要通过小鼠足垫技术进行验证，并且对于研究皮肤涂片样本中麻风分枝杆菌的存活情况、治疗效果以及其他分枝杆菌的存在，也应进行更广泛的探索。
