Abstract
Carbapenem-resistant A. baumannii (CRAB) represents a public health threat increasing worldwide. We assess the suitability of a loop-mediated isothermal amplification (LAMP) method for on-site screening of CRAB in a hospital facility. A set of six primers were designed for recognizing eight distinct sequences on six targets: blaOXA-23-like, blaOXA-24-like, blaOXA-51-like, blaOXA-58-like, blaIMP, and blaVIM. A LAMP method was developed, optimized and evaluated for the identification of CRAB in thirty-three environmental samples from an outbreak in an Intensive Care Unit (ICU) facility. The sensitivity of the LAMP assay for the detection of A. baumannii was ten-fold higher than the PCR assay (1.0 ng.µL-1). The LAMP assays showed a higher detection rate for CRAB samples and robust diagnosis performance in comparison to a conventional PCR, with clinical sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of 100% for blaOXA-23-like, blaOXA-51-like and blaVIM. The developed LAMP assays are powerful tools that can be useful in on-site screening of CRAB causing local outbreaks in clinics and hospitals facilities where costs and equipment restraints are imperative.
Highlights
Carbapenem-resistant A. baumannii (CRAB) represents a public health threat increasing worldwide
This study developed loop-mediated isothermal amplification (LAMP) detection assays for CRAB by targeting blaOXA23-like, blaOXA-24-like, blaOXA-51-like, blaOXA-58-like, blaIMP and blaVIM genes and evaluated the assays performance with environmental samples from an outbreak in an Intensive Care Unit (ICU) facility
Clinical isolates collected from Mexican hospitals [39,40]; including A. baumannii 9001, A. baumannii 8400, A. baumannii 7811, P. aeruginosa 4899 and P. aeruginosa 6102, known to produce OXA-23-like, OXA-24-like, OXA-58-like, IMP and Verona Imipenemase (VIM), respectively, were used as positive controls to determine the appropriate conditions for LAMP
Summary
Carbapenem-resistant A. baumannii (CRAB) represents a public health threat increasing worldwide. Conclusions: The developed LAMP assays are powerful tools that can be useful in on-site screening of CRAB causing local outbreaks in clinics and hospitals facilities where costs and equipment restraints are imperative. Acinetobacter baumannii environmental widespread and propensity to cause large and multifacility nosocomial outbreaks have been demonstrated clearly [5,6,7]. It can survive and persist in dry conditions for extended time periods in the patients’ environment as a result of its capacity to form biofilms on biotic and abiotic surfaces [8,9,10]. A. baumannii has been recovered from various sites including furniture, patient care items and hospital equipment [9,11,12].
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