Specific amplification of Aspergillus fumigatus DNA by polymerase chain reaction.
Specific amplification of Aspergillus fumigatus DNA by polymerase chain reaction.
复制标题
通过聚合酶链式反应特异性扩增烟曲霉 DNA。
DOI：
10.1006/mcpr.1993.1016
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发表时间：
1993
影响因子：
3.3
通讯作者：
V. Kurup
中科院分区：
文献类型：
--
作者：
L. V. Reddy;Anoopa Kumar;V. Kurup
我们已经开发了一种特异性和敏感性的方法来检测人类病原体烟曲霉菌的聚合酶链反应（PCR）的目的是检测生物体在外周血和尿液中，可以通过非侵入性的程序。根据已发表的A.烟曲霉Asp f1和烟曲霉的核毒素丝裂霉素和局限菌素。restrictus和A.阿格特斯结果表明，所设计的引物能特异扩增出A.烟曲霉A. restrictus而不是A.阿格特斯此外，没有人DNA或A. Flavus、黄毛菊A.尼日尔、黑曲霉A. fischeri、青霉菌属、白色念珠菌和卡氏肺孢子虫。PCR检测A.烟曲霉DNA在溴化乙锭凝胶上约为20 pg，通过使用32P标记的内部寡核苷酸的Southern分析为0.6 pg。对13例疑似侵袭性曲霉病（IA）患者的尿液标本进行初步分析，2例PCR阳性，其中1例死于IA伴脑损害。目前正在进一步分析IA的尿液和血液标本，以确定PCR与传统抗原检测的比较效用。
We have developed a specific and sensitive method to detect the human pathogen Aspergillus fumigatus by polymerase chain reaction (PCR) with an objective of detecting the organism in peripheral blood and urine which can be obtained by non-invasive procedures. A pair of oligonucleotide primers for PCR were designed based on the published partial protein sequences of an 18 KD IgE-binding protein of A. fumigatus Asp f1 and the ribotoxins mitogillin and restrictocin of A. restrictus, and alpha-sarcin of A. giganteus. The primers were specific in amplifying an expected 315 bp region of the homologous genes in A. fumigatus and A. restrictus but not in A. giganteus. Also, there was no amplification of human DNA or DNA of A. flavus, A. niger, A. fischeri, Penicillium sp., Candida albicans and Pneumocystis carinii. The sensitivity of the PCR detection of A. fumigatus DNA is about 20 pg on an ethidium bromide gel and 0.6 pg by Southern analysis using a 32P-labelled internal oligonucleotide. In preliminary analysis of 13 urine specimens of patients suspected of invasive aspergillosis (IA), two were PCR positive, one of whom died of IA with brain lesion. Further analyses of both urine and blood specimens of IA are in progress to determine the comparative utility of PCR over the conventional antigen tests.