聚合酶链反应扩增和测序在非流行地区旧世界利什曼原虫感染诊断和分型中的前瞻性价值
Prospective value of PCR amplification and sequencing for diagnosis and typing of old world Leishmania infections in an area of nonendemicity.
作者信息
Gangneux Jean-Pierre, Menotti Jean, Lorenzo Frédéric, Sarfati Claudine, Blanche Hélène, Bui Hung, Pratlong Francine, Garin Yves-Jean-François, Derouin Francis
机构信息
Laboratoire de Parasitologie-Mycologie, Centre Hospitalier Universitaire Lariboisière-Saint-Louis, 75010 Paris, France.
出版信息
J Clin Microbiol. 2003 Apr;41(4):1419-22. doi: 10.1128/JCM.41.4.1419-1422.2003.
Abstract
We assessed the prospective value of PCR amplification of a repetitive sequence from Leishmania nuclear DNA and sequencing for the diagnosis and typing of Old World Leishmania infection in an area of nonendemicity. During this 42-month study, 29 of 168 consecutive samples were examined and classified as positive for Leishmania by direct examination and/or in vitro culture. This molecular approach showed excellent sensitivity (97%) and specificity (100%) compared to direct examination (86 and 100%, respectively) and in vitro culture (72 and 100%, respectively). Isoenzymatic and molecular typing allowed similar identification for 12 samples. Besides, PCR and subsequent sequencing of DNA products permitted the species identification of 14 samples for which parasite culture remained negative or did not allow isoenzymatic characterization, indicating the complementarity of parasitological and molecular tools.
摘要
我们评估了从利什曼原虫核DNA中扩增一个重复序列并进行测序用于非流行地区旧世界利什曼原虫感染诊断和分型的前瞻性价值。在这项为期42个月的研究中，对168个连续样本中的29个进行了检测，并通过直接检查和/或体外培养将其分类为利什曼原虫阳性。与直接检查（分别为86%和100%）和体外培养（分别为72%和100%）相比，这种分子方法显示出优异的敏感性（97%）和特异性（100%）。同工酶和分子分型对12个样本的鉴定结果相似。此外，PCR及后续DNA产物测序使14个样本得以进行物种鉴定，这些样本的寄生虫培养结果仍为阴性或无法进行同工酶特征分析，这表明寄生虫学和分子工具具有互补性。
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