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Comparison of the FXG™: RESP (Asp+) real-time PCR assay with direct immunofluorescence and calcofluor white staining for the detection of Pneumocystis jirovecii in respiratory specimens

, , , , , , , & show all
Pages 324-327 | Received 16 Mar 2011, Accepted 17 Jun 2011, Published online: 23 Aug 2011
 

Abstract

We compared the FXG™: RESP (Asp +) real-time PCR assay (Myconostica Ltd) with two microscopic staining methods (direct immunofluorescence [IFA] and calcofluor white) for the detection of Pneumocystis jirovecii in 411 respiratory specimens submitted for P. jirovecii examination. We considered the specimen to be microscopically positive if the organism could be visualized through the use of either IFA or calcofluor white. A second, published real-time PCR assay targeting the cdc2 gene of P. jirovecii was used to adjudicate those specimens that were microscopically negative but Myconostica PCR positive. The Myconostica PCR positive samples were deemed to be true positives if they were concordant with microscopically positive results or if they were positive by the second PCR assay. As a result, the Myconostica PCR assay was found to be more sensitive than the two microscopy methods in detecting P. jirovecii (10.5% true positivity rate by PCR, 8.0% by immunofluorescence, and 7.1% by calcofluor white). The Myconostica PCR assay showed 93.5% sensitivity, 95.1% specificity, 70.5% positive predictive value, and 99.1% negative predictive value. Its high negative predictive value suggests a role of the Myconostica PCR assay in ruling out Pneumocystis pneumonia.

Acknowledgements

We would like to thank Don Martin, Daisy Yang, Tiffany Fung and Aimin Li for their tremendous technical help with this study.

Declaration of interest: Dr John Thornback is an employee of Myconostica Ltd. The other authors report no conflicts of interest. The authors alone are responsible for the content and writing of the paper.

This paper was first published online on Early Online on 22 August 2011.

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