Loudon K W, Burnie J P, Coke A P, Matthews R C
Department of Medical Microbiology, University of Manchester Medical School, United Kingdom.
J Clin Microbiol. 1993 May;31(5):1117-21. doi: 10.1128/jcm.31.5.1117-1121.1993.
A new method for fingerprinting Aspergillus fumigatus by random amplification of polymorphic DNA (RAPD) by using single primers with arbitrary sequences is described. Five primers were examined with 19 isolates from six patients with aspergilloma as well as with A. fumigatus NCPF 2109. Two of the primers (GCT GGT GG and GCG CAC GG, 5' to 3') gave adequate discrimination between isolates, generating five and six types, respectively. Combination of the results obtained with each of these two primers generated 12 types. This compares very favorably with immunoblot fingerprinting and XbaI-generated restriction fragment length polymorphisms on the same isolates. Typeability and reproducibility were good with RAPD, and RAPD was less labor-intensive than immunoblot fingerprinting. RAPD typing results suggested that aspergillomas sometimes contain isolates of more than one type.
描述了一种通过使用具有任意序列的单引物对烟曲霉进行随机扩增多态性DNA(RAPD)指纹图谱分析的新方法。用来自6例曲菌球患者的19株分离菌以及烟曲霉NCPF 2109对5种引物进行了检测。其中两种引物(GCT GGT GG和GCG CAC GG,5'至3')能够对分离菌进行充分区分,分别产生5种和6种类型。将这两种引物各自获得的结果相结合产生了12种类型。这与对相同分离菌进行免疫印迹指纹图谱分析和XbaI酶切产生的限制性片段长度多态性分析相比,具有很大优势。RAPD的分型能力和可重复性良好,并且与免疫印迹指纹图谱分析相比,RAPD的劳动强度较小。RAPD分型结果表明,曲菌球有时包含不止一种类型的分离菌。