Nazzal Duà, Yasin Salem, Abu-Elteen Khaled
Biological Sciences Department, Hashemite University, Al-Zarqa, Jordan.
New Microbiol. 2005 Jul;28(3):245-50.
The rapid detection and identification of Candida species in clinical laboratories are extremely important for the management of patients with hematogenous candidosis. Currently available culture and biochemical methods for detection and identification of Candida species are time-consuming. This study describes the use of a simple and rapid PCR method using species-specific oligonucleotides for the detection of clinical isolates of Candida species. These species-specific oligonucleotides are complementary to unique sequences within the intergenic transcribed spacer 2, located in between the 5.8S and 28S ribosomal DNA, and generated DNA fragments by both the conventional and hemi-nested PCR reactions. Conventional PCR produced a single DNA fragment of variable size in all isolates, while the hemi-nested PCR produced two discrete DNA fragments, both with the expected sizes of 111bp/57bp (C. albicans), 84bp/42bp (C. glabrata), 94bp/45bp (C. krusei) and 95bp/49bp (C. parapsilosis). In conclusion, the PCR-based method described in this study is fast and specific for the identification of clinically important Candida species.
临床实验室中念珠菌属的快速检测和鉴定对于血源性念珠菌病患者的管理极为重要。目前用于念珠菌属检测和鉴定的培养及生化方法耗时较长。本研究描述了一种使用物种特异性寡核苷酸的简单快速PCR方法,用于检测念珠菌属的临床分离株。这些物种特异性寡核苷酸与位于5.8S和28S核糖体DNA之间的基因间隔转录间隔区2内的独特序列互补,并通过常规PCR反应和半巢式PCR反应产生DNA片段。常规PCR在所有分离株中产生大小可变的单一DNA片段,而半巢式PCR产生两个离散的DNA片段,其大小均符合预期,分别为白色念珠菌的111bp/57bp、光滑念珠菌的84bp/42bp、克柔念珠菌的94bp/45bp和近平滑念珠菌的95bp/49bp。总之,本研究中描述的基于PCR的方法对于鉴定临床上重要的念珠菌属快速且特异。