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Comparative Study
. 2019 Mar;39(2):176-182.
doi: 10.3343/alm.2019.39.2.176.

Comparative Evaluation Between the RealStar Pneumocystis jirovecii PCR Kit and the AmpliSens Pneumocystis jirovecii (carinii)-FRT PCR Kit for Detecting P. jirovecii in Non-HIV Immunocompromised Patients

Hee Jae Huh #  1 , Kyoung Ree Lim #  2 , Chang Seok Ki  3 , Kyungmin Huh  2 , Hyang Jin Shim  4 , Dong Joon Song  1 , Yae Jean Kim  5 , Doo Ryeon Chung  2   6 , Nam Yong Lee  7
Affiliations
Comparative Study

Comparative Evaluation Between the RealStar Pneumocystis jirovecii PCR Kit and the AmpliSens Pneumocystis jirovecii (carinii)-FRT PCR Kit for Detecting P. jirovecii in Non-HIV Immunocompromised Patients

Hee Jae Huh et al. Ann Lab Med. 2019 Mar.

Abstract

Background: Real-time PCR is more sensitive than microscopic examination for detecting Pneumocystis jirovecii. We compared the performance of two assays for detecting P. jirovecii DNA: the RealStar Pneumocystis jirovecii PCR Kit 1.0 CE (Altona Diagnostics, Hamburg, Germany) and the AmpliSens Pneumocystis jirovecii (carinii)-FRT PCR kit (InterLabService Ltd., Moscow, Russia).

Methods: We used 159 samples from the lower respiratory tract (112 bronchoalveolar lavage [BAL] fluid, 37 sputum, and 10 endotracheal aspirate [ETA] samples) of non-HIV immunocompromised patients. Nested PCR and sequencing were used to resolve discordant results. The performance of the two assays was evaluated according to clinical categories (clinical Pneumocystis pneumonia [PCP], possible PCP, or unlikely PCP) based on clinical and radiological observations.

Results: The positive and negative percent agreement values were 100% (95% confidence interval [CI], 85.4-100%) and 96.6% (95% CI, 90.9-98.9%), respectively, and kappa was 0.92 (95% CI, 0.84-0.99). P. jirovecii DNA load was significantly higher in the clinical PCP group than in the other groups (P<0.05). When stratified by sample type, the positive rate for BAL fluids from the clinical PCP group was 100% using either assay, whereas the positive rate for sputum/ETA samples was only 20%.

Conclusions: The two assays showed similar diagnostic performance and detected low P. jirovecii burden in BAL fluids. Both assays may be useful as routine methods for detecting P. jirovecii DNA in a clinical laboratory setting, though their results should be interpreted considering sample type.

Keywords: AmpliSens Pneumocystis jirovecii (carinii)-FRT PCR; Performance; Pneumocystis jirovecii; Pneumocystis pneumonia; Real-time PCR; RealStar Pneumocystis jirovecii PCR.

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Conflict of interest statement

No potential conflicts of interest relevant to this article were reported.

Figures

Fig. 1
Fig. 1. Scatter plot showing a strong correlation between the quantitative results (log copies/mL) obtained using the RealStar assay and the Ct values obtained using the AmpliSens assay (r=−0.94; 95% confidence interval, −0.97 to −0.88).
Abbreviation: Ct, Cycle threshold.
Fig. 2
Fig. 2. Distribution of P. jirovecii DNA load according to clinical probability of PCP. (A) Distribution of quantitative results (log copies/mL) obtained using the RealStar assay in total samples; (B) Distribution of the Ct values obtained using the AmpliSens assay in total samples; (C) Distribution of quantitative results (log copies/mL) obtained by the RealStar assay in BAL samples; (D) Distribution of Ct values obtained by the AmpliSens assay in BAL samples. Median log copies/mL or Ct values and IQRs are indicated (central horizontal lines, median; outer horizontal lines, 25–95% IQR).
Abbreviations: Ct, Cycle threshold; BAL, bronchoalveolar lavage; IQR, interquartile range; PCP, Pneumocystis pneumonia.
See this image and copyright information in PMC

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