DEVELOPMENT AND CLINICAL VALIDATION OF AN INTEGRATED RAPID POINT OF CARE WORKFLOW FOR DETECTION OF BLACTX-M-15 AND AAC (6')-IB-CR ANTIMICROBIAL RESISTANCE GENES IN URINARY TRACT INFECTION URINE AT RESOURCE LIMITED HOSPITALS AND LABORATORIES.

Abstract

Background: The Rapid detection of antimicrobial resistance (AMR) is essential for periodically diagnosing for antimicrobial analysis, yet the conventional antimicrobial susceptibility testing (AST) method still requires a minimum of 24–72 hours. This study developed and validate an integrated point-of-care real-time PCR workflow for rapid detection of the blaCTX-M-15 and aac (6')-Ib-cr resistance genes extracted from urine specimens. Methods: A total of 150 urine samples from adults with suspected urinary tract infections were analyzed using HiPurA Genomic DNA extraction Kits followed by real-time PCR. This diagnostic is a simple molecular workflow which is rapid, simple, and cost effectiveness and can be used in low resource laboratories in the detection of antimicrobial resistance. It has been was evaluated against the phenotypic AST and other already existed molecular methods, Results: The complete sample to result molecular workflow was completed within approximately 60 minutes. The workflow was demonstrated at high diagnostic performance for blaCTX-M-15 with the diagnostic threshold of 94.74% sensitivity and a specificity of 90.91% followed by the other resistance gene which aac (6')-Ib-cr which was having a diagnostic performance of sensitivity 92.63% and specificity of 92.73% with the reference to the Antimicrobial Susceptibility Testing method. Conclusions: This study involves basically present a rapid molecular and PCR workflow which is directly extracted from urine sample targeting blaCTX-M-15 and aac (6′)-Ib-cr within a time space of 60minutes