Evaluation of the PneumoGenius® PCR assay for the diagnosis ofPneumocystispneumonia and the detection ofPneumocystisdihydropteroate synthase mutations in respiratory samples

Author:

Guegan Hélène12ORCID,Roojee Maël2,Le Gal Solène34,Artus Mathilde34,Nevez Gilles34ORCID,Gangneux Jean-Pierre12,Robert-Gangneux Florence12

Affiliation:

1. Université de Rennes, Institut de Recherche en Santé, Environnement et travail , Inserm UMR_S 1085, Rennes , France

2. Laboratoire de Parasitologie-Mycologie , Centre Hospitalier Universitaire de Rennes, Rennes , France

3. Laboratoire de Parasitologie-Mycologie, Centre Hospitalier Universitaire de Brest , Brest , France

4. Infections Respiratoires Fongiques, Université de Brest/Angers , Brest/Angers , France

Abstract

AbstractPneumocystis pneumonia (PCP) is the most frequent fungal opportunistic infection defining AIDS in HIV-infected patients, and is of growing importance in HIV-negative patients. In this latter category of patients, the diagnosis mainly relies on real-time polymerase chain reaction (qPCR) detection of Pneumocystis jirovecii (Pj) on respiratory samples. The PneumoGenius® kit (PathoNostics) allows the simultaneous detection of Pj mitochondrial large subunit (mtLSU) and dihydropteroate synthase (DHPS) polymorphisms, which could be of interest to anticipate therapeutic failure. This study aimed at evaluating its clinical performance on 251 respiratory specimens (239 patients), (i) for P. jirovecii detection in clinical samples, and (ii) for DHPS polymorphisms detection in circulating strains.Patients were classified according to modified European Organization for Research and Treatment of Cancer/Invasive Fungal Infections Cooperative Group and the National Institute of Allergy and Infectious Diseases Mycoses Study Group (EORTC/MSG) criteria, as having proven PCP (n = 62), probable PCP (n = 87), Pneumocystis colonization (n = 37), and no PCP (n = 53).Compared with in-house qPCR, the sensitivity of PneumoGenius® assay for P. jirovecii detection reached 91.9% (182/198), the specificity was excellent (100%, 53/53) and the global concordance was 93.6% (235/253). A total of four diagnoses of proven/probable PCP were missed by the PneumoGenius® assay, reaching a 97.5% sensitivity (157/161) in this sub-group. The 12 other ‘false–negative’ results were obtained in patients diagnosed as colonized using the in-house PCR. DHPS genotyping was successful for 147/182 samples with PneumoGenius® and revealed dhps mutation in 8 samples, which were all confirmed by sequencing.In conclusion, PneumoGenius® assay missed the detection of low-burden PCP. This lower sensitivity for PCP diagnosis can be balanced by a higher specificity (P. jirovecii colonization less frequently detected) and the efficient detection of DHPS hot spot mutations.

Funder

Institut de Parasitologie de l'Ouest

Publisher

Oxford University Press (OUP)

Subject

Infectious Diseases,General Medicine

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