中华皮肤科杂志 ›› 2002, Vol. 35 ›› Issue (2): 128-130.

• 论著 • 上一篇    下一篇

应用种特异性DNA探针原位杂交技术检测系统性曲霉感染

马蕾1, 李若瑜2   

  1. 1. 北京中日友好医院皮肤科 100029;
    2. 北京大学第一医院皮肤科真菌和真菌病研究中心
  • 收稿日期:2001-09-18 出版日期:2002-04-15 发布日期:2002-04-15
  • 基金资助:
    教育部优秀跨世纪人材基金及教育部骨干教师计划资助项目

In Situ Hybridization Technique for Detecting Systemic Aspergillus Infection Using A Species Specific DNA Probe

MA Lei1, LI Ruoyu2   

  1. 1. Deptof Dermatology &Research Center for Medical Mycology, First Hospital, Peking University,Beijing 100034;
    2. Department of Dermatology,China Japan Friendship Hospital, Beijing 100029, China
  • Received:2001-09-18 Online:2002-04-15 Published:2002-04-15

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摘要： 目的 探讨应用原位杂交技术(ISH)特异性检测系统性曲霉感染临床标本的可行性。方法 以对烟曲霉高度特异的碱性蛋白酶基因片段作为种特异性探针,聚合酶链反应(PCR)扩增制备,并以地高辛标记,与16例福尔马林固定、石蜡包埋疑为系统性曲霉感染的临床组织标本中的靶DNA杂交。结果 16例可疑曲霉感染标本13例阳性,其他真菌感染组织均为阴性。结论 该技术可以敏感、特异地检测出组织中曲霉的感染。由于此探针是烟曲霉特异的,并与黄曲霉有一定的同源性,推论应用该DNA探针可以筛选出临床常见的这两种曲霉的感染,适用于临床曲霉感染的检测。

关键词: 曲霉病, 原位杂交, DNA探针

Abstract: Objectives To evaluate the applicability of in situ hybridization (ISH) technique in the specific diagnosis of clinical systemic Aspergillus infection. Methods Sixteen formalin fixed and paraffin embeded tissue specimens from patients with suspected Aspergillus spp. infection were studied by an ISH technique with a DNA probe representing fragment of alkaline proteinase gene, which was synthesized and digoxigenin labeled, by means of polymerase chain reaction (PCR). Results Positive staining was obtained in 13 tissue specimens from 16 patients with suspected systemic Aspergillus infection, but in none of the tissues infected with other fungi. Conclusions The ISH is a sensitive, specific technique for detecting systemic Aspergillus infection. As the DNA probe is known specific for Aspergillus fumigatus and has low homology with Aspergillus flavus, the ISH technique with this probe could differentiate the two common species from other Aspergillus spp. or from non Aspergillus fungi in tissue specimens. It could be applicable to clinical molecular diagnosis of Aspergillus infection.

Key words: DNA probes, In situ hybridization, Aspergillosis