Department of Dermatology, The Second Affiliated Hospital, Sun Yat-Sen University, Guangzhou, China.
Mycoses. 2011 Nov;54(6):e751-9. doi: 10.1111/j.1439-0507.2011.02017.x. Epub 2011 Sep 19.
Penicillium marneffei is the aetiological agent of a severe systemic disease in immunocompromised hosts in Southeast Asia. In the present study, we evaluated an identification method based on rolling circle amplification (RCA) enabling rapid and specific detection of single nucleotide differences. Three padlock probes were designed on the basis of the internal transcribed spacers 1 and 2 (ITS) of the rRNA operon. One of these (PmPL1) allowed specific amplification of P. marneffei DNA within one working day using a newly conceived protocol, while no cross-reactivity was observed with other fungi including related biverticillate penicillia. Amplification products can be detected by electrophoresis on agarose gel. The method provides a powerful tool for a rapid specific identification of P. marneffei in the clinical laboratory and has potential for ecological studies.
马尔尼菲青霉是免疫功能低下宿主中一种严重全身性疾病的病原体。在本研究中，我们评估了一种基于滚环扩增（RCA）的鉴定方法，该方法能够快速、特异地检测单核苷酸差异。根据 rRNA 操纵子的内部转录间隔区 1 和 2（ITS）设计了三个套锁探针。其中一个（PmPL1）使用新构思的方案，在一个工作日内允许马尔尼菲青霉 DNA 的特异性扩增，而与其他真菌（包括相关的双轮青霉）没有交叉反应。扩增产物可通过琼脂糖凝胶电泳检测。该方法为临床实验室中快速、特异性鉴定马尔尼菲青霉提供了有力工具，并且具有生态研究的潜力。
