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印度黑热病一株旧临床分离株的分子鉴定

Molecular identification of an old clinical isolate of Indian Kala-azar.

作者信息

Manna Madhumita, Khanra Supriya

机构信息

Department of Zoology, Barasat Govt. College, 10, K.N.C Road, Kolkata, 700124 India.

出版信息

J Parasit Dis. 2017 Jun;41(2):336-344. doi: 10.1007/s12639-016-0799-9. Epub 2016 Jun 4.

Abstract

Molecular characterization is an important task for species identification of the isolates belonging to different species. Clinical symptoms, tissue tropism, vector preference, reservoir and geographical distribution may act as distinguishing parameters but not always decisive. On the other hand, modern taxonomic tools employed to divulge characteristics of the genome or protein molecules of the parasite would be convincing and for sp., they include nuclear and kDNA buoyant density, multi locus enzyme electrophoresis (MLEE), RAPD, RFLP or use of monoclonal antibodies etc. In the present study, we intend to establish the identification of an old clinical isolate of Indian Kala-azar, familiarly known as 'UR6' by MLEE, RAPD, RFLP and species specific monoclonal antibodies. UR6 has been isolated from a confirmed Kala-azar patient admitted in Calcutta School of Tropical Medicine, Kolkata in 1978. From then it is being regularly used for various scientific studies by the Research Group of India and abroad. The isozyme profile of UR6 showed similar electrophoretic mobility that of WHO reference strain for K27. Similar findings were obtained in the RAPD and RFLP assays. Screening with species specific monoclonal antibodies showed its strong reactivity towards . The Jaccard's Similarity Indices were calculated.

摘要

分子特征分析是鉴定不同物种分离株的一项重要任务。临床症状、组织嗜性、媒介偏好、宿主和地理分布可能是区分参数,但并非总是决定性的。另一方面,用于揭示寄生虫基因组或蛋白质分子特征的现代分类工具更具说服力,对于利什曼原虫属物种,这些工具包括核DNA和动基体DNA浮力密度、多位点酶电泳(MLEE)、随机扩增多态性DNA(RAPD)、限制性片段长度多态性(RFLP)或使用单克隆抗体等。在本研究中,我们打算通过MLEE、RAPD、RFLP和物种特异性单克隆抗体来鉴定一株印度黑热病的老临床分离株,俗称“UR6”。UR6于1978年从加尔各答热带医学院收治的一名确诊黑热病患者中分离得到。从那时起,印度国内外的研究小组一直在定期将其用于各种科学研究。UR6的同工酶谱显示出与K27的世界卫生组织参考菌株相似的电泳迁移率。在RAPD和RFLP分析中也得到了类似的结果。用物种特异性单克隆抗体进行筛选显示其对杜氏利什曼原虫有强烈反应。计算了杰卡德相似性指数。

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