在生物样本中使用65 kDa甘露糖蛋白基因引物进行白色念珠菌的实时PCR鉴定。
Use of 65 kDa mannoprotein gene primers in Real Time PCR identification of Candida albicans in biological samples.
作者信息
Arancia Silvia, Carattoli Alessandra, La Valle Roberto, Cassone Antonio, De Bernardis Flavia
机构信息
Department of Infectious, Parasitic and Immune-mediated Diseases, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161, Rome, Italy.
出版信息
Mol Cell Probes. 2006 Oct;20(5):263-8. doi: 10.1016/j.mcp.2006.01.006. Epub 2006 Feb 23.
A method for the detection and quantification of Candida albicans in biological samples (blood, urine and serum) was developed with the use of Real-Time PCR utilizing CaMP65-specific primers. Two different systems were used for the detection in the LightCycler platform (Roche): the SYBR green fluorescent dye with melting peak analysis and the 5'nuclease fluorescent-probe detection. The amplification was highly specific for C. albicans, providing no cross-reaction on genomic DNA extracted from other Candida species or Aspergillus. The sensitivity in simulated biological samples was especially high (1 genome) when applied to sera and urine, and in blood samples the limit of detection was higher by ten-fold. Finally, the real-time PCR was employed in order to detect and quantify C. albicans in the sera from patients with invasive candidiasis.
利用针对CaMP65的特异性引物，通过实时聚合酶链反应（Real-Time PCR）开发了一种用于检测和定量生物样本（血液、尿液和血清）中白色念珠菌的方法。在LightCycler平台（罗氏公司）上使用了两种不同的系统进行检测：带有熔解峰分析的SYBR绿色荧光染料和5'核酸酶荧光探针检测。该扩增对白色念珠菌具有高度特异性，对从其他念珠菌属物种或曲霉菌中提取的基因组DNA无交叉反应。当应用于血清和尿液时，在模拟生物样本中的灵敏度特别高（1个基因组），而在血液样本中检测限高出十倍。最后，采用实时PCR来检测和定量侵袭性念珠菌病患者血清中的白色念珠菌。
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