A Real-Time PCR Assay Based on 5.8S rRNA Gene (5.8S rDNA) for Rapid Detection of Candida from Whole Blood Samples

被引:11
作者
Guo, Yi [1 ]
Yang, Jing-xian [1 ]
Liang, Guo-wei [1 ]
机构
[1] Peking Univ, Dept Clin Lab, Aerosp Sch Clin Med, 15 Yuquan Rd, Beijing 100049, Peoples R China
关键词
Candida; Real-time PCR; 5.8S rRNA gene (5.8S rDNA); Whole blood samples; Bloodstream infections (BSIs); PROSPECTIVE NATIONWIDE SURVEILLANCE; STREAM INFECTIONS; FUNGAL PATHOGENS; IDENTIFICATION; MULTIPLEX; CULTURE; DNA; EPIDEMIOLOGY; DIAGNOSIS; GLABRATA;
D O I
10.1007/s11046-015-9977-z
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
The prevalence of Candida in bloodstream infections (BSIs) has increased. To date, the identification of Candida in BSIs still mainly relies on blood culture and serological tests, but they have various limitations. Therefore, a real-time PCR assay for the detection of Candida from whole blood is presented. The unique primers/probe system was designed on 5.8S rRNA gene (5.8S rDNA) of Candida genus. The analytical sensitivity was determined by numbers of positive PCRs in 12 repetitions. At the concentration of 10(1) CFU/ml blood, positive PCR rates of 100 % were obtained for C. albicans, C. parapsilosis, C. tropicalis, and C. krusei. The detection rate for C. glabrata was 75 % at 10(1) CFU/ml blood. The reaction specificity was 100 % when evaluating the assay using DNA samples from clinical isolates and human blood. The maximum CVs of intra-assay and inter-assay for the detection limit were 1.22 and 2.22 %, respectively. To assess the clinical applicability, 328 blood samples from 82 patients were prospectively tested and real-time PCR results were compared with results from blood culture. Diagnostic sensitivity of the PCR was 100 % using as gold standard blood culture, and specificity was 98.4 %. Our data suggest that the developed assay can be used in clinical laboratories as an accurate and rapid screening test for the Candida from whole blood. Although further evaluation is warranted, our assay holds promise for earlier diagnosis of candidemia.
引用
收藏
页码:405 / 413
页数:9
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