用于快速诊断溃疡分枝杆菌感染的聚合酶链反应检测方法的开发。
Development of a PCR assay for rapid diagnosis of Mycobacterium ulcerans infection.
作者信息
Ross B C, Marino L, Oppedisano F, Edwards R, Robins-Browne R M, Johnson P D
机构信息
Department of Microbiology and Infectious Disease, Royal Children's Hospital, Parkville, Australia.
出版信息
J Clin Microbiol. 1997 Jul;35(7):1696-700. doi: 10.1128/jcm.35.7.1696-1700.1997.
The diagnosis of Mycobacterium ulcerans infection is hampered by the slow growth of the bacterium in culture, resulting in a delay of several months before a specific diagnosis can be obtained. In addition, M. ulcerans cannot be isolated from water even when there is convincing epidemiological evidence implicating this as the source of infection. The aim of the present study was to develop a PCR assay to circumvent the problems of delayed diagnosis and insensitivity of standard bacterial culture for M. ulcerans. For the PCR, we isolated an M. ulcerans-specific DNA fragment, 1,109 bp long, which is repeated at least 50 times throughout the genome. Use of this sequence as a target for PCR allowed us to detect as few as 2 molecules of genomic DNA in vitro. The PCR was used to detect M. ulcerans DNA in fresh tissue and paraffin-embedded sections from all seven patients with culture-confirmed cases of infection.
溃疡分枝杆菌感染的诊断因该细菌在培养物中生长缓慢而受到阻碍，导致在获得特异性诊断之前会延迟数月。此外，即使有确凿的流行病学证据表明水是感染源，也无法从水中分离出溃疡分枝杆菌。本研究的目的是开发一种聚合酶链反应（PCR）检测方法，以解决溃疡分枝杆菌标准细菌培养诊断延迟和不敏感的问题。对于PCR，我们分离出一个1109 bp长的溃疡分枝杆菌特异性DNA片段，该片段在整个基因组中至少重复50次。以该序列作为PCR的靶标，使我们能够在体外检测到低至2个基因组DNA分子。PCR被用于检测来自所有7例经培养确诊感染病例的新鲜组织和石蜡包埋切片中的溃疡分枝杆菌DNA。
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