Dunyach Catherine, Bertout Sébastien, Phelipeau Cécile, Drakulovski Pascal, Reynes Jacques, Mallié Michele
Laboratoire de Parasitologie et de Mycologie Médicale, UMR 145 (IRD-UM1) <<VIH-SIDA et maladies associées>>, Faculté de Pharmacie, BP 14491, 15, 34093 Montpellier Cedex 5, France.
Diagn Microbiol Infect Dis. 2008 Mar;60(3):263-71. doi: 10.1016/j.diagmicrobio.2007.09.014. Epub 2007 Nov 26.
The aim of this work was to develop LightCycler real-time polymerase chain reaction method to allow rapid detection and identification of Candida spp. in human serum with panfungal primers (internal transcribed spacer [ITS] and L18). Melting-curve analysis of the ITS sequences showed that each amplicon presented a specific melting point and enabled identification of 5 Candida spp. After parameters optimization, 58 sera were preliminary analyzed from 23 patients. For L18 primers, the LightCycler system enabled detection of DNA in 92% of patients with positive blood culture. These primers were not able to differentiate between species of Candida. By using ITS primers, the LightCycler system enabled detection of DNA in sera from 76.9% of patients with positive blood culture. With ITS primers, the species responsible for the infection was identified for 11 patients. These data revealed the LightCycler as a potential tool for early detection and identification of Candida.
这项工作的目的是开发一种LightCycler实时聚合酶链反应方法，以便使用泛真菌引物（内转录间隔区[ITS]和L18）快速检测和鉴定人血清中的念珠菌属。ITS序列的熔解曲线分析表明，每个扩增子都呈现出特定的熔点，能够鉴定5种念珠菌属。参数优化后，对23例患者的58份血清进行了初步分析。对于L18引物，LightCycler系统能够在92%血培养阳性的患者中检测到DNA。这些引物无法区分念珠菌的种类。使用ITS引物时，LightCycler系统能够在76.9%血培养阳性的患者血清中检测到DNA。使用ITS引物时，为11例患者鉴定出了引起感染的菌种。这些数据表明，LightCycler是早期检测和鉴定念珠菌的一种潜在工具。
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