Abstract
A multiplex TaqMan real-time PCR to detect carbapenem-hydrolysing class D b-lactamases (blaOXA-23-like, blaOXA-24/40-like, blaOXA-51-like and blaOXA-58-like genes) was developed and evaluated for early detection of imipenem (IMP) resistance in clinically significant Acinetobacter baumannii isolates. Well-characterized strains of A. baumannii were used as positive controls and non-Acinetobacter strains were used to assess specificity. Analytical sensitivity was quantified by comparison with the number of bacterial c.f.u. Forty of 46 (87 %) clinically significant and IMPresistant A. baumannii isolates were positive for the blaOXA-23-like gene, and one isolate (2 %) was positive for the blaOXA-58-like gene. The blaOXA-24/40-like gene was not detected in any of the 46 IMP-resistant strains and the blaOXA-51-like gene was identified in both IMP-resistant and non-resistant A. baumannii. All 11 non-Acinetobacter bacteria produced a negative result for each of the four blaOXAgenes. This assay was able to detect as few as 10 c.f.u. per assay. This real-time PCR method demonstrated rapid detection of OXA-like carbapenem resistance in A. baumannii in comparison with phenotypic susceptibility testing methodology. This method could be adapted to a multiplexed single reaction for rapid detection of genes associated with carbapenem resistance in A. baumannii and potentially other clinically significant multidrugresistant Gram-negative bacteria.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 1532-1537 |
| Number of pages | 6 |
| Journal | Journal of Medical Microbiology |
| Volume | 61 |
| Issue number | PART 11 |
| DOIs | |
| State | Published - Nov 1 2012 |
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All Science Journal Classification (ASJC) codes
- Microbiology
- Microbiology (medical)
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Development and validation of a multiplex TaqMan real-time PCR for rapid detection of genes encoding four types of class D carbapenemase in Acinetobacter baumannii. / Huang, Xiao Zhe; Cash, Dana M.; Chahine, Mohamad A.; Nikolich, Mikeljon P.; Craft, David W.
In: Journal of Medical Microbiology, Vol. 61, No. PART 11, 01.11.2012, p. 1532-1537.Research output: Contribution to journal › Article
TY - JOUR
T1 - Development and validation of a multiplex TaqMan real-time PCR for rapid detection of genes encoding four types of class D carbapenemase in Acinetobacter baumannii
AU - Huang, Xiao Zhe
AU - Cash, Dana M.
AU - Chahine, Mohamad A.
AU - Nikolich, Mikeljon P.
AU - Craft, David W.
PY - 2012/11/1
Y1 - 2012/11/1
N2 - A multiplex TaqMan real-time PCR to detect carbapenem-hydrolysing class D b-lactamases (blaOXA-23-like, blaOXA-24/40-like, blaOXA-51-like and blaOXA-58-like genes) was developed and evaluated for early detection of imipenem (IMP) resistance in clinically significant Acinetobacter baumannii isolates. Well-characterized strains of A. baumannii were used as positive controls and non-Acinetobacter strains were used to assess specificity. Analytical sensitivity was quantified by comparison with the number of bacterial c.f.u. Forty of 46 (87 %) clinically significant and IMPresistant A. baumannii isolates were positive for the blaOXA-23-like gene, and one isolate (2 %) was positive for the blaOXA-58-like gene. The blaOXA-24/40-like gene was not detected in any of the 46 IMP-resistant strains and the blaOXA-51-like gene was identified in both IMP-resistant and non-resistant A. baumannii. All 11 non-Acinetobacter bacteria produced a negative result for each of the four blaOXAgenes. This assay was able to detect as few as 10 c.f.u. per assay. This real-time PCR method demonstrated rapid detection of OXA-like carbapenem resistance in A. baumannii in comparison with phenotypic susceptibility testing methodology. This method could be adapted to a multiplexed single reaction for rapid detection of genes associated with carbapenem resistance in A. baumannii and potentially other clinically significant multidrugresistant Gram-negative bacteria.
AB - A multiplex TaqMan real-time PCR to detect carbapenem-hydrolysing class D b-lactamases (blaOXA-23-like, blaOXA-24/40-like, blaOXA-51-like and blaOXA-58-like genes) was developed and evaluated for early detection of imipenem (IMP) resistance in clinically significant Acinetobacter baumannii isolates. Well-characterized strains of A. baumannii were used as positive controls and non-Acinetobacter strains were used to assess specificity. Analytical sensitivity was quantified by comparison with the number of bacterial c.f.u. Forty of 46 (87 %) clinically significant and IMPresistant A. baumannii isolates were positive for the blaOXA-23-like gene, and one isolate (2 %) was positive for the blaOXA-58-like gene. The blaOXA-24/40-like gene was not detected in any of the 46 IMP-resistant strains and the blaOXA-51-like gene was identified in both IMP-resistant and non-resistant A. baumannii. All 11 non-Acinetobacter bacteria produced a negative result for each of the four blaOXAgenes. This assay was able to detect as few as 10 c.f.u. per assay. This real-time PCR method demonstrated rapid detection of OXA-like carbapenem resistance in A. baumannii in comparison with phenotypic susceptibility testing methodology. This method could be adapted to a multiplexed single reaction for rapid detection of genes associated with carbapenem resistance in A. baumannii and potentially other clinically significant multidrugresistant Gram-negative bacteria.
UR - http://www.scopus.com/inward/record.url?scp=84867521075&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84867521075&partnerID=8YFLogxK
U2 - 10.1099/jmm.0.045823-0
DO - 10.1099/jmm.0.045823-0
M3 - Article
C2 - 22878252
AN - SCOPUS:84867521075
VL - 61
SP - 1532
EP - 1537
JO - Journal of Medical Microbiology
JF - Journal of Medical Microbiology
SN - 0022-2615
IS - PART 11
ER -