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Development and evaluation of a real-time PCR assay for detection of Pneumocystis jiroveci.
Transpl Infect Dis. 2007 Sep; 9(3):196-202.TI

Abstract

Pneumocystis jiroveci is an important agent of pneumonia in immunocompromised hosts. Usually, this pathogen is detected by Giemsa or direct fluorescence stains of bronchoalveolar lavage (BAL) fluids. Microscopic methods, however, have 2 disadvantages. P. jiroveci is not stable outside the human body, which means that slow sample transport might result in false-negative results. Additionally, exact quantification, which is needed for therapy monitoring, is not possible. In this study, we developed a real-time polymerase chain reaction assay for the LightCycler. Two Pneumocystis-specific TaqMan systems, one based on the sequence of the 5.8S ribosomal gene and another one targeting the dihydrofolate reductase gene were evaluated. Additionally, the amount of human DNA in the sample was measured by a TaqMan assay based on the human albumin gene, allowing assessment of sample quality and quantification normalized on sample concentration. For clinical evaluation, 69 BAL specimens from 26 positive patients as well as 60 negative controls were tested. Both systems were able to detect all proven cases of Pneumocystis pneumonia. Differentiation of carriage, asymptomatic reactivation, and clinical infection as well as normalized quantification by calculating the ratio of Pneumocystis DNA to human DNA are discussed.

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Authors+Show Affiliations

Bandt D
Institut für Virologie, Medizinisch-Theoretisches Zentrum, Dresden, Germany. Dirk.Bandt@mailbox.tu-dresden.de
Monecke S
No affiliation info available

MeSH

Bronchoalveolar Lavage FluidDNA, BacterialHumansMicroscopyPneumocystis cariniiPneumonia, PneumocystisPolymerase Chain ReactionRNA, Ribosomal, 5.8SSensitivity and SpecificityTetrahydrofolate Dehydrogenase

Pub Type(s)

Journal Article

Language

eng

PubMed ID

17605743

Citation

Bandt, D, and S Monecke. "Development and Evaluation of a Real-time PCR Assay for Detection of Pneumocystis Jiroveci." Transplant Infectious Disease : an Official Journal of the Transplantation Society, vol. 9, no. 3, 2007, pp. 196-202.
Bandt D, Monecke S. Development and evaluation of a real-time PCR assay for detection of Pneumocystis jiroveci. Transpl Infect Dis. 2007;9(3):196-202.
Bandt, D., & Monecke, S. (2007). Development and evaluation of a real-time PCR assay for detection of Pneumocystis jiroveci. Transplant Infectious Disease : an Official Journal of the Transplantation Society, 9(3), 196-202.
Bandt D, Monecke S. Development and Evaluation of a Real-time PCR Assay for Detection of Pneumocystis Jiroveci. Transpl Infect Dis. 2007;9(3):196-202. PubMed PMID: 17605743.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Development and evaluation of a real-time PCR assay for detection of Pneumocystis jiroveci. AU - Bandt,D, AU - Monecke,S, Y1 - 2007/07/01/ PY - 2007/7/4/pubmed PY - 2007/12/12/medline PY - 2007/7/4/entrez SP - 196 EP - 202 JF - Transplant infectious disease : an official journal of the Transplantation Society JO - Transpl Infect Dis VL - 9 IS - 3 N2 - Pneumocystis jiroveci is an important agent of pneumonia in immunocompromised hosts. Usually, this pathogen is detected by Giemsa or direct fluorescence stains of bronchoalveolar lavage (BAL) fluids. Microscopic methods, however, have 2 disadvantages. P. jiroveci is not stable outside the human body, which means that slow sample transport might result in false-negative results. Additionally, exact quantification, which is needed for therapy monitoring, is not possible. In this study, we developed a real-time polymerase chain reaction assay for the LightCycler. Two Pneumocystis-specific TaqMan systems, one based on the sequence of the 5.8S ribosomal gene and another one targeting the dihydrofolate reductase gene were evaluated. Additionally, the amount of human DNA in the sample was measured by a TaqMan assay based on the human albumin gene, allowing assessment of sample quality and quantification normalized on sample concentration. For clinical evaluation, 69 BAL specimens from 26 positive patients as well as 60 negative controls were tested. Both systems were able to detect all proven cases of Pneumocystis pneumonia. Differentiation of carriage, asymptomatic reactivation, and clinical infection as well as normalized quantification by calculating the ratio of Pneumocystis DNA to human DNA are discussed. SN - 1398-2273 UR - https://www.unboundmedicine.com/medline/citation/17605743/Development_and_evaluation_of_a_real_time_PCR_assay_for_detection_of_Pneumocystis_jiroveci_ L2 - https://doi.org/10.1111/j.1399-3062.2007.00246.x DB - PRIME DP - Unbound Medicine ER -