Diagnostic significance of nested polymerase chain reaction for sensitive detection of Pneumocystis jirovecii in respiratory clinical specimens.

Full Abstract

A total of 327 clinical specimens, including both invasive and noninvasive samples, obtained from 275 patients with various types of underlying immunocompromised conditions and a clinical suspicion of Pneumocystis pneumonia (PCP) were subjected to 2 different nested polymerase chain reaction (PCR) assays. The target genes used for nested PCR were mitochondrial large subunit ribosomal RNA (mtLSU rRNA) and internal transcribed spacer (ITS) region. The results were compared with a single-round PCR targeting major surface glycoprotein (MSG) gene. Amplification was successful in 16% of cases by mtLSU rRNA nested PCR, in 14.5% by ITS nested PCR, and in 10.9% by MSG PCR. The nested mtLSU rRNA PCR was found to be more sensitive (100% sensitive and 98.7% specific) and useful in detecting PCP for its use in routine diagnosis in our settings. Thus, this assay may be quite useful in the identification of patients who are in the early stage of Pneumocystis jirovecii infection with an organism load that could not be easily detected by the single-step PCR.

Author information

Author/s: Gupta, Rashmi (R); Mirdha, Bijay Ranjan (BR); Guleria, Randeep (R); Kumar, Lalit (L); Samantaray, Jyotish Chandra (JC); Agarwal, Sanjay Kumar (SK); Kabra, Sushil Kumar (SK); Luthra, Kalpana (K);

Affiliation: Department of Microbiology, All Indian Institute of Medical Sciences, Ansari Nagar, New Delhi 110029, India.

Journal and publication information

Publication Type: Comparative Study; Evaluation Studies; Journal Article; Research Support, Non-U.S. Gov't

Journal: Diagnostic microbiology and infectious disease (Diagn Microbiol Infect Dis), published in United States. (Language: eng)

Reference: 2009-Aug; vol 64 (issue 4) : pp 381-8

Dates: Created 2009/07/27; Completed 2009/09/21;

PMID: 19631091, status: MEDLINE (last retrieval date: 9/21/2009, IMS Date: )

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MeSH headings (categories)

This article was linked to the MESH Headings shown below.

DNA Primers - genetics
DNA, Fungal - genetics
DNA, Ribosomal Spacer - genetics
Humans
Molecular Sequence Data
Pneumocystis Infections - diagnosis; microbiology
Pneumocystis jirovecii - genetics; isolation & purification

Pneumocystis jirovecii - genetics; isolation & purification
Polymerase Chain Reaction - methods
RNA - genetics
RNA, Ribosomal - genetics
Sensitivity and Specificity
Sequence Analysis, DNA
Sputum - microbiology

Associated Chemicals: DNA Primers (0) ; DNA, Fungal (0) ; DNA, Ribosomal Spacer (0) ; RNA, Ribosomal (0) ; RNA, mitochondrial (0) ; RNA (63231-63-0)

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