Use of a multiplex PCR-based reverse line blot (mPCR/RLB) hybridisation assay for the rapid identification of bacterial pathogens.
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 x 10(2) CFU/mL for Streptococcus pneumoniae and 6 x 10(2) 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.
Wang
Y
Y
Beijing Children's Hospital Affiliated to Capital Medical University, Beijing, China.
Kong
F
F
Gilbert
G L
GL
Brown
M
M
Gao
W
W
Yu
S
S
Yang
Y
Y
D001419:Bacteria / Q000145:classification* / Q000235:genetics; D001424:Bacterial Infections / Q000175:diagnosis; D001431:Bacteriological Techniques / Q000379:methods*; D001483:Base Sequence; D015139:Blotting, Southern / Q000379:methods*; D027081:DNA Gyrase / Q000235:genetics; D004269:DNA, Bacterial / Q000032:analysis; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013045:Species Specificity
D004269:DNA, Bacterial; D027081:DNA Gyrase