Tags

Type your tag names separated by a space and hit enter

Development and Evaluation of a Fully Automated Molecular Assay Targeting the Mitochondrial Small Subunit rRNA Gene for the Detection of Pneumocystis jirovecii in Bronchoalveolar Lavage Fluid Specimens.
J Mol Diagn. 2020 12; 22(12):1482-1493.JM

Abstract

The fungal pathogen Pneumocystis jirovecii causes Pneumocystis pneumonia. Although the mitochondrial large subunit rRNA gene (mtLSU) is commonly used as a PCR target, a mitochondrial small subunit rRNA gene (mtSSU)-targeted MultiCode PCR assay was developed on the fully automated ARIES platform for detection of P. jirovecii in bronchoalveolar lavage fluid specimens in 2.5 hours. The assay showed a limit of detection of 800 copies/mL (approximately equal to 22 organisms/mL), with no cross-reactivity with other respiratory pathogens. Compared with the reference Pneumocystis-specific direct fluorescent antibody assay (DFA) and mtLSU-targeted PCR assay, the new assay demonstrated sensitivity of 96.9% (31/32) and specificity of 94.6% (139/147) in detecting P. jirovecii in 180 clinical bronchoalveolar lavage fluid specimens. This assay was concordant with all DFA-positive samples and all but one mtLSU PCR-positive sample, and detected eight positive samples that were negative by DFA and mtLSU PCR. Receiver operating characteristic curve analysis revealed an area under the curve of 0.98 and a threshold cycle (CT) cutoff of 39.1 with sensitivity of 90.9% and specificity of 99.3%. The detection of 39.1 <CT < 40.0 indicates the presence of a low load of the organism and needs further determination of either colonization or probable/possible Pneumocystis pneumonia. Overall, the new assay demonstrates excellent analytical and clinical performance and may be more sensitive than mtLSU PCR target for the detection of P. jirovecii.

Links

Publisher Full Text (DOI)

Authors+Show Affiliations

Liu B
Division of Medical Microbiology, Department of Pathology, Johns Hopkins University School of Medicine, Baltimore, Maryland.
Totten M
Division of Medical Microbiology, Department of Pathology, Johns Hopkins University School of Medicine, Baltimore, Maryland.
Nematollahi S
Division of Infectious Diseases, Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, Maryland.
Datta K
Division of Infectious Diseases, Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, Maryland.
Memon W
Microbiology Laboratory, Johns Hopkins Hospital, Baltimore, Maryland.
Marimuthu S
Division of Infectious Diseases, Department of Medicine, University of Louisville School of Medicine, Louisville, Kentucky.
Wolf LA
Division of Infectious Diseases, Department of Medicine, University of Louisville School of Medicine, Louisville, Kentucky.
Carroll KC
Division of Medical Microbiology, Department of Pathology, Johns Hopkins University School of Medicine, Baltimore, Maryland; Microbiology Laboratory, Johns Hopkins Hospital, Baltimore, Maryland.
Zhang SX
Division of Medical Microbiology, Department of Pathology, Johns Hopkins University School of Medicine, Baltimore, Maryland; Microbiology Laboratory, Johns Hopkins Hospital, Baltimore, Maryland. Electronic address: szhang28@jhmi.edu.

MeSH

AdolescentAdultAgedAged, 80 and overBronchoalveolar Lavage FluidChildChild, PreschoolFemaleFluorescent Antibody Technique, DirectGenes, rRNAHumansInfantLimit of DetectionMaleMiddle AgedMitochondrial RibosomesMolecular Diagnostic TechniquesPneumocystis cariniiPneumonia, PneumocystisRNA, RibosomalReal-Time Polymerase Chain ReactionRetrospective StudiesSensitivity and SpecificityYoung Adult

Pub Type(s)

Evaluation Study
Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

33069878

Citation

Liu, Baoming, et al. "Development and Evaluation of a Fully Automated Molecular Assay Targeting the Mitochondrial Small Subunit rRNA Gene for the Detection of Pneumocystis Jirovecii in Bronchoalveolar Lavage Fluid Specimens." The Journal of Molecular Diagnostics : JMD, vol. 22, no. 12, 2020, pp. 1482-1493.
Liu B, Totten M, Nematollahi S, et al. Development and Evaluation of a Fully Automated Molecular Assay Targeting the Mitochondrial Small Subunit rRNA Gene for the Detection of Pneumocystis jirovecii in Bronchoalveolar Lavage Fluid Specimens. J Mol Diagn. 2020;22(12):1482-1493.
Liu, B., Totten, M., Nematollahi, S., Datta, K., Memon, W., Marimuthu, S., Wolf, L. A., Carroll, K. C., & Zhang, S. X. (2020). Development and Evaluation of a Fully Automated Molecular Assay Targeting the Mitochondrial Small Subunit rRNA Gene for the Detection of Pneumocystis jirovecii in Bronchoalveolar Lavage Fluid Specimens. The Journal of Molecular Diagnostics : JMD, 22(12), 1482-1493. https://doi.org/10.1016/j.jmoldx.2020.10.003
Liu B, et al. Development and Evaluation of a Fully Automated Molecular Assay Targeting the Mitochondrial Small Subunit rRNA Gene for the Detection of Pneumocystis Jirovecii in Bronchoalveolar Lavage Fluid Specimens. J Mol Diagn. 2020;22(12):1482-1493. PubMed PMID: 33069878.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Development and Evaluation of a Fully Automated Molecular Assay Targeting the Mitochondrial Small Subunit rRNA Gene for the Detection of Pneumocystis jirovecii in Bronchoalveolar Lavage Fluid Specimens. AU - Liu,Baoming, AU - Totten,Marissa, AU - Nematollahi,Saman, AU - Datta,Kausik, AU - Memon,Warda, AU - Marimuthu,Subathra, AU - Wolf,Leslie A, AU - Carroll,Karen C, AU - Zhang,Sean X, Y1 - 2020/10/15/ PY - 2020/05/18/received PY - 2020/09/10/revised PY - 2020/10/01/accepted PY - 2020/10/19/pubmed PY - 2021/11/9/medline PY - 2020/10/18/entrez SP - 1482 EP - 1493 JF - The Journal of molecular diagnostics : JMD JO - J Mol Diagn VL - 22 IS - 12 N2 - The fungal pathogen Pneumocystis jirovecii causes Pneumocystis pneumonia. Although the mitochondrial large subunit rRNA gene (mtLSU) is commonly used as a PCR target, a mitochondrial small subunit rRNA gene (mtSSU)-targeted MultiCode PCR assay was developed on the fully automated ARIES platform for detection of P. jirovecii in bronchoalveolar lavage fluid specimens in 2.5 hours. The assay showed a limit of detection of 800 copies/mL (approximately equal to 22 organisms/mL), with no cross-reactivity with other respiratory pathogens. Compared with the reference Pneumocystis-specific direct fluorescent antibody assay (DFA) and mtLSU-targeted PCR assay, the new assay demonstrated sensitivity of 96.9% (31/32) and specificity of 94.6% (139/147) in detecting P. jirovecii in 180 clinical bronchoalveolar lavage fluid specimens. This assay was concordant with all DFA-positive samples and all but one mtLSU PCR-positive sample, and detected eight positive samples that were negative by DFA and mtLSU PCR. Receiver operating characteristic curve analysis revealed an area under the curve of 0.98 and a threshold cycle (CT) cutoff of 39.1 with sensitivity of 90.9% and specificity of 99.3%. The detection of 39.1 <CT < 40.0 indicates the presence of a low load of the organism and needs further determination of either colonization or probable/possible Pneumocystis pneumonia. Overall, the new assay demonstrates excellent analytical and clinical performance and may be more sensitive than mtLSU PCR target for the detection of P. jirovecii. SN - 1943-7811 UR - https://www.unboundmedicine.com/medline/citation/33069878/Development_and_Evaluation_of_a_Fully_Automated_Molecular_Assay_Targeting_the_Mitochondrial_Small_Subunit_rRNA_Gene_for_the_Detection_of_Pneumocystis_jirovecii_in_Bronchoalveolar_Lavage_Fluid_Specimens_ DB - PRIME DP - Unbound Medicine ER -
Grapherence [↓5]

Related Citations

More