학술논문
Use of a multiplex PCR-based reverse line blot (mPCR/RLB) hybridisation assay for the rapid identification of bacterial pathogens.
Document Type
Article
Source
Subject
*PATHOGENIC microorganisms
*NUCLEIC acid hybridization
*PATHOGENIC bacteria
*MEDICAL bacteriology
*BACTERIAL diseases
*DNA
*GENES
*STREPTOCOCCUS
*ENTEROBACTERIACEAE
*FUNGUS-bacterium relationships
*STREPTOCOCCUS pneumoniae
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Language
ISSN
1198-743X
Abstract
The aim of this study was to develop a sensitive and reliable method for the molecular identification of pathogenic bacteria. A multiplex PCR-based reverse line blot (mPCR/RLB) hybridisation assay was developed and evaluated for the rapid identification of 24 systemic and respiratory bacterial pathogens in routine diagnosis. All species-specific probes designed for the RLB hybridised with amplified DNA only from the corresponding species. Sensitivity limits of the mPCR/RLB assay varied among the 24 target organisms from 0.05 pg to 0.5 ng of genomic DNA. The sensitivity of the assay was 2 × 102 CFU/mL for Streptococcus pneumoniae and 6 × 102 CFU/mL for Escherichia coli. The specificity of each probe was tested against 24 species. There were no cross-reactions among any of the 43 probes. The mPCR/RLB assay appeared to be a useful alternative tool for the molecular identification of common pathogens. [ABSTRACT FROM AUTHOR]