Increased sensitivity of a new commercial reverse transcriptase-quantitative PCR for the detection of Pneumocystis jirovecii in respiratory specimens

Sarah Dellière (1, 2) , Samia Hamane (1) , Nesrine Aissaoui (1) , Maud Gits-Muselli (1, 2) , Stéphane Bretagne (1) , Alexandre Alanio (1, 2, 3)

1 Groupe Hospitalier Saint Louis - Lariboisière - Fernand Widal [Paris]
2 Mycologie moléculaire - Molecular Mycology
3 CNRMA - Centre National de Référence Mycoses Invasives et Antifongiques - National Reference Center Invasive Mycoses & Antifungals

Optimal sensitivity to detect low Pneumocystis loads is of importance to take individual and collective measures to avoid evolution towards Pneumocystis pneumonia and outbreaks in immunocompromised patients. This study compares two qPCR procedures, a new automated RTqPCR using the GeneLEAD VIII extractor/thermocycler (GLVIII; ∼2.2 hrs workflow) and a previously validated in-house qPCR assays (IH; ∼5 hrs workflow) both targeting mtSSU and mtLSU for detecting P. jirovecii in 213 respiratory samples. GLVIII was found to be more sensitive than IH, detecting 8 more specimens. Bland-Altman analysis between the two procedures showed a Cq bias of 1.17 ± 0.07 in favor of GLVIII. The fungus Pneumocystis needs to be detected early in respiratory samples to prevent pneumonia in immunocompromised hosts. We evaluated a new commercial RTqPCR on 213 respiratory samples to detect Pneumocystis and found it more sensitive and faster than our routine sensitive in-house qPCR assay.

Keywords: Pneumocystis jirovecii; real time PCR; diagnosis; Pneumocystis pneumonia; mitochondria

Domains: Mycology; Systematics, Phylogenetics and taxonomy; Infectious diseases

Sarah Dellière, Samia Hamane, Nesrine Aissaoui, Maud Gits-Muselli, Stéphane Bretagne, et al.. Increased sensitivity of a new commercial reverse transcriptase-quantitative PCR for the detection of Pneumocystis jirovecii in respiratory specimens. Medical Mycology, 2021, pp.myab029. ⟨10.1093/mmy/myab029⟩. ⟨pasteur-03226313⟩