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

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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 101 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 101 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.

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Acknowledgments

The authors thank Baskova L, et al. (authors of “The Pan-AC assay: a single-reaction real-time PCR test for quantitative detection of a broad range of Aspergillus and Candida species”) for providing the description of procedure of specificity assessment. The study was funded by a college research grant from the Aerospace Center Hospital (YN201322).

Funding

The study was funded by a college research grant from the Aerospace Center Hospital (YN201322).

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Authors and Affiliations

  1. Department of Clinical Laboratory, Peking University Aerospace School of Clinical Medicine, No. 15 Yuquan Road, Haidian District, Beijing, 100049, China

    Yi Guo, Jing-xian Yang & Guo-wei Liang

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  1. Yi Guo
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  2. Jing-xian Yang
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  3. Guo-wei Liang
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Correspondence to Guo-wei Liang.

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The authors declare that they have no conflict of interest.

Human and Animal Rights

The research involved human participants who donated their blood for preparing simulated candidemia samples and assessing clinical application for the PCR method.

Ethical Approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of Aerospace Center Hospital committee and with the 1964 Declaration of Helsinki and its later amendments or comparable ethical standards.

Informed Consent

Informed consent was obtained from all individual participants included in the study.

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Guo, Y., Yang, Jx. & Liang, Gw. A Real-Time PCR Assay Based on 5.8S rRNA Gene (5.8S rDNA) for Rapid Detection of Candida from Whole Blood Samples. Mycopathologia 181, 405–413 (2016). https://doi.org/10.1007/s11046-015-9977-z

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  • DOI: https://doi.org/10.1007/s11046-015-9977-z

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