通过两步聚合酶链反应检测和鉴定结核分枝杆菌、牛分枝杆菌/卡介苗和鸟分枝杆菌。与使用A60抗原的酶联免疫吸附测定法的比较。
Detection and identification of Mycobacterium tuberculosis, M. bovis/BCG, and M. avium by two-step polymerase chain reaction. Comparison with ELISA using A60 antigen.
作者信息
Bollet C, De Lamballerie X, Zandotti C, Vignoli C, Gevaudan M J, De Micco P
机构信息
Laboratoire de Microbiologie et d'Hygiène Hospitalière, Hôpital Salvator, Marseille, France.
出版信息
Microbiologica. 1992 Oct;15(4):345-9.
We propose a rapid two-step PCR to amplify a 767-bp sequence present in the gene coding for the 65-kD antigen of mycobacteria. The high G+C content (80%) permitted annealing to occur at 70 degrees C, enhancing the specificity. The amplified fragment contains a restriction site for differentiation between M. tuberculosis, M. bovis/BCG, and M. avium. Complete diagnosis can be achieved in less than four hours without labelled probe or nucleic acid transfer.
我们提出一种快速两步聚合酶链反应（PCR）方法，用于扩增分枝杆菌65-kD抗原编码基因中一段767碱基对的序列。其高鸟嘌呤-胞嘧啶（G+C）含量（80%）使得退火反应能在70℃进行，从而提高了特异性。扩增片段包含一个限制性酶切位点，可用于区分结核分枝杆菌、牛分枝杆菌/卡介苗和鸟分枝杆菌。无需标记探针或核酸转移，在不到四小时内即可完成诊断。
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