title: A Real-Time PCR Assay Based on 5.8S rRNA Gene (5.8S rDNA) for Rapid Detection of Candida from Whole Blood Samples.
abstract: 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.
authors.affiliation: Department of Clinical Laboratory, Peking University Aerospace School of Clinical Medicine, No. 15 Yuquan Road, Haidian District, Beijing, 100049, China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Guo
authors.affiliation: Department of Clinical Laboratory, Peking University Aerospace School of Clinical Medicine, No. 15 Yuquan Road, Haidian District, Beijing, 100049, China.
authors.forename: Jing-Xian
authors.initials: JX
authors.lastname: Yang
authors.affiliation: Department of Clinical Laboratory, Peking University Aerospace School of Clinical Medicine, No. 15 Yuquan Road, Haidian District, Beijing, 100049, China. aerospace_guoyi@hotmail.com.
authors.forename: Guo-Wei
authors.initials: GW
authors.lastname: Liang
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D001769:Blood / Q000382:microbiology*; D002175:Candida / Q000235:genetics / Q000302:isolation & purification*; D058387:Candidemia / Q000175:diagnosis*; D017931:DNA Primers / Q000235:genetics; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D012340:RNA, Ribosomal, 5.8S / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D017931:DNA Primers; D012340:RNA, Ribosomal, 5.8S
keywords: 5.8S rRNA gene (5.8S rDNA); Bloodstream infections (BSIs); Candida; Real-time PCR; Whole blood samples