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Cost-effective Method for Microbial Source Tracking Using Specific Human and Animal Viruses
Published on: December 3, 2011
A Cost-Effective Method for Identifying Enterobacterales with OXA-181
Gisele Peirano1,2, Yasufumi Matsumura3, Diego Nobrega4
1Division of Microbiology, Alberta Public Laboratories, Cummings School of Medicine, University of Calgary, Calgary, Alberta, Canada.
Abstract:
OXA-181 is the second most common global OXA-48-like carbapenemase and is endemic in the Indian subcontinent. Molecular studies have shown that Enterobacterales with OXA-181 are often introduced into regions of nonendemicity. Distinguishing OXA-181 from other OXA-48-like enzymes often requires sequencing, which is rather expensive and time-consuming. A specific PCR (i.e., OXA181PCR) for the detection of blaOXA-181 was validated using a global collection (n = 315) of bacteria with well-characterized carbapenemases and showed 100% sensitivity and specificity (95% confidence interval [CI], 94.1 to 100 and 98.6 to 100, respectively) for detecting bacteria with OXA-181. The OXA181PCR subsequently gave positive results on 58/160 (36%) Enterobacterales with OXA-48-like carbapenemases from the 2015 INFORM surveillance program. The blaOXA-181-positive Enterobacterales were present in 9 countries spanning 5 continents, illustrating the global distribution of OXA-181. This methodology can easily be incorporated into molecular surveillance programs to provide accurate information about the prevalence of OXA-181. A loop-mediated isothermal amplification (LAMP)-OXA48 assay overall performed well for detecting OXA-48-like enzymes but showed poor specificity due to false-positive results with non-OXA carbapenemases.
Insights
A new PCR test accurately detects OXA-181, a common carbapenemase found globally. This rapid molecular method aids surveillance of OXA-181 producing Enterobacterales, crucial for infection control.
Area of Science:
- Microbiology
- Molecular Biology
- Infectious Diseases
Background:
- OXA-181 is a prevalent carbapenemase, particularly in the Indian subcontinent, often spreading to non-endemic regions.
- Distinguishing OXA-181 from similar enzymes typically requires costly and time-consuming sequencing.
- Effective surveillance requires rapid and accurate detection methods for OXA-181.
Purpose of the Study:
- To validate a specific Polymerase Chain Reaction (PCR) assay, OXA181PCR, for the accurate detection of the blaOXA-181 gene.
- To assess the prevalence and global distribution of OXA-181 producing Enterobacterales using the developed PCR method.
- To evaluate the utility of OXA181PCR in molecular surveillance programs.
Main Methods:
- Validation of OXA181PCR using a global collection of 315 bacteria with characterized carbapenemases.
- Application of OXA181PCR to 160 Enterobacterales isolates from the 2015 INFORM surveillance program.
- Comparison of OXA181PCR performance with a loop-mediated isothermal amplification (LAMP)-OXA48 assay.
Main Results:
- OXA181PCR demonstrated 100% sensitivity and specificity for detecting OXA-181.
- The assay identified OXA-181 in 36% (58/160) of Enterobacterales from the INFORM surveillance program.
- OXA-181 positive isolates were found in 9 countries across 5 continents, indicating widespread dissemination.
Conclusions:
- OXA181PCR is a highly sensitive and specific molecular tool for detecting OXA-181.
- The method facilitates accurate and efficient surveillance of OXA-181 producing bacteria globally.
- Incorporating OXA181PCR into surveillance programs can provide crucial data on the prevalence and spread of this important carbapenemase.
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