Development of TaqMan probe-based real-time PCR method for erm(A),erm(B), and erm(C), rapid detection of macrolide-lincosamide-streptogramin B resistance genes, from clinical isolates.
To achieve more accurate and rapid detection of macrolidelincosamide- streptogramin B resistance genes, erm(A), erm(B), and erm(C), we developed a TaqMan probe-based real-time PCR (Q-PCR) method and compared it with conventional PCR (C-PCR), which is the most widely using erm gene identification method. The detection limit of Q-PCR was 5 fg of genomic DNA or 5-8 CFU of bacterial cells of Staphylococcus aureus. The utilization of Q-PCR might shorten the time to erm detection from 3-4 h to about 50 min. These data indicated that Q-PCR assay appears to be not only highly sensitive and specific, but also the most rapid diagnostic method. Therefore, the appropriate application of the Q-PCR assay will permit rapid and accurate identification of erm genes from clinical and other samples.
Jung
Jae-Hyuk
JH
College of Pharmacy, Sahmyook University, 139-742 Seoul, Korea.
Yoon
Eun-Jeong
EJ
Choi
Eung-Chil
EC
Choi
Sung-Sook
SS
D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001426:Bacterial Proteins / Q000032:analysis / Q000235:genetics*; D003428:Cross Infection / Q000382:microbiology*; D017931:DNA Primers; D015342:DNA Probes; D024901:Drug Resistance, Multiple, Bacterial / Q000235:genetics; D006094:Gram-Positive Bacteria / Q000187:drug effects / Q000235:genetics* / Q000302:isolation & purification; D016908:Gram-Positive Bacterial Infections / Q000382:microbiology*; D006801:Humans; D055231:Lincosamides / Q000494:pharmacology; D018942:Macrolides / Q000494:pharmacology; D008780:Methyltransferases / Q000032:analysis / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D025381:Streptogramin B / Q000494:pharmacology; D013997:Time Factors
D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D017931:DNA Primers; D015342:DNA Probes; D055231:Lincosamides; D018942:Macrolides; D025381:Streptogramin B; D008780:Methyltransferases; C431709:ErmA protein, Bacteria; C035181:rRNA (adenosine-O-2'-)methyltransferase