Loading…

Quantitative reverse transcription-PCR assay for the rapid detection of methicillin-resistant Staphylococcus aureus

To evaluate a new quantitative reverse transcription-PCR (qRT-PCR) assay for the rapid detection of methicillin-resistant Staphylococcus aureus (MRSA). Primers for Staphylococcus-specific regions of 16S rRNA gene, spa gene and mecA gene were newly designed. RNAs extracted from broth-cultured strains...

Full description

Saved in:
Bibliographic Details
Published in:Journal of applied microbiology 2010-03, Vol.108 (3), p.779-788
Main Authors: Wada, M, Lkhagvadorj, E, Bian, L, Wang, C, Chiba, Y, Nagata, S, Shimizu, T, Yamashiro, Y, Asahara, T, Nomoto, K
Format: Article
Language:English
Subjects:
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:To evaluate a new quantitative reverse transcription-PCR (qRT-PCR) assay for the rapid detection of methicillin-resistant Staphylococcus aureus (MRSA). Primers for Staphylococcus-specific regions of 16S rRNA gene, spa gene and mecA gene were newly designed. RNAs extracted from broth-cultured strains were tested by qRT-PCR targeting each primer, and the bacterial counts obtained correlated well with those counted by the plating method with detection limits of 10⁰, 10¹ and 10² CFU. The qRT-PCR assay targeting the 16S rRNA was 6430-fold or more sensitive than qPCR assay. All Staph. aureus strains tested were detected and none of the other Staphylococcus species and genus strains tested cross-reacted with the assay targeting the spa gene. All MRSAs tested were detected by the assay targeting the mecA gene. Clinical samples, faecal material and bronchial washout solutions were tested by our assay, and MRSAs were detected with a high sensitivity within 6 h. Our qRT-PCR assay targeting three new primers to the target genes is a rapid and sensitive tool for the detection of MRSA directly from clinical samples. Because of its sensitivity and rapidity, our qRT-PCR assay is considered to be a valuable tool for clinical management.
ISSN:1364-5072
1365-2672
DOI:10.1111/j.1365-2672.2009.04476.x