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. 2019 Sep 17;5(3):86.
doi: 10.3390/jof5030086.

Diagnostic Performance of a Novel Multiplex PCR Assay for Candidemia among ICU Patients

Stefan Fuchs  1 , Cornelia Lass-Flörl  1 , Wilfried Posch  2
Affiliations

Diagnostic Performance of a Novel Multiplex PCR Assay for Candidemia among ICU Patients

Stefan Fuchs et al. J Fungi (Basel). .

Abstract

Candidemia poses a major threat to ICU patients and is routinely diagnosed by blood culture, which is known for its low sensitivity and long turnaround times. We compared the performance of a novel, Candida-specific multiplex real-time PCR assay (Fungiplex® Candida IVD Real-Time PCR Kit) with blood culture and another established diagnostic real-time PCR assay (LightCycler SeptiFast Test) with respect to Candida detection from whole blood samples. Clinical samples from 58 patients were analyzed by standard blood culture (BC) and simultaneously tested with the Fungiplex Candida PCR (FP) and the SeptiFast test (SF) for molecular detection of Candida spp. Compared to BC, the FP test showed high diagnostic power, with a sensitivity of 100% and a specificity of 94.1%. Overall diagnostic accuracy reached 94.6%. Using SF, we found a sensitivity of 60%, a specificity of 96.1%, and an overall diagnostic accuracy of 92.9%. The Fungiplex Candida PCR has shown good sensitivity and specificity on clinical samples of high-risk patients for direct detection of Candida species in whole blood samples. Together with conventional diagnostics (BC and antigen testing), this new multiplex PCR assay may contribute to a rapid and accurate diagnosis of candidiasis.

Keywords: Candida PCR; candidemia; molecular diagnostics; yeast multiplex PCR.

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Conflict of interest statement

The authors declare no conflict of interest. In the past 5 years, W.P. has received grant support from the OeNB Anniversary Fund. C.L.-F. and W.P. have received grant support from the Austrian Science Fund (FWF). C.L.-F. has been supported by Astellas Pharma, Gilead Sciences, Pfizer, Schering Plough, and Merck Sharp and Dohme. She has been an advisor/consultant to Gilead Sciences, Merck Sharp and Dohme, Pfizer, and Schering Plough. She has received honoraria for talks and travel costs from Gilead Sciences, Merck Sharp and Dohme, Pfizer, Astellas Pharma, and Schering Plough.

Figures

Figure 1
Figure 1
Diagnostic workflow including identification of Candida spp. in blood. Blood samples are sent for both blood culture and PCR-based diagnostics - Fungiplex (FP) and SeptiFast (SF). Ten milliliters of whole blood are injected into aerobic and anaerobic blood culture bottles. The SF test and FP PCR are run using the same DNA extract from 1 mL of EDTA blood. Positive blood cultures are subcultivated and analyzed using a MALDI-TOF system for species identification (ID). Results are reported as positive (+) including the identified species, negative (-) or, if PCR controls failed, as inhibited (inh). The time-to-result (TTR) varies between 1–7 days for BC (including ID) and 3–5 h for the molecular tests.
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