The qPCR Assay for Detecting The Presence and Relative Abundance of Pseudomonas aerugionosa and Antibiotic Resistance Gene aadA2 in Hospital Wastewater of National Reference Hospital Dr. Cipto Mangunkusumo (RSCM)

Abstract

Antimicrobial resistance is one of the top 10 global health threats. The hospital wastewater (HWW) potentially becomes the reservoir and dissemination of antibiotic resistance gene (ARG) and bacterial pathogens. In Indonesia, the protocol to monitor the ARGs form HWW has not been established. This study aimed to detect the presence and find the relative abundance of P. aeruginosa and aadA2 genes from Dr. RSUPN. Cipto Mangungkusumo (RSCM) inlet and outlet wastewater through qPCR assay. The primers used were supported by Resistomap. The study revealed that the qPCR assay was able to detect the Ct value of P. aeruginosa and aadA2. The aadA2 gene was found in all waste water samples, meanwhile P. aeruginosa was only found in some of inlet samples. aadA2 had the highest relative abundance and this gene’s mobility uses plasmids and integrons that potentially enhance the acquired antimicrobial resistance (AMR) mechanism. This study implicated that qPCR assay was capable to detect pathogenic bacteria and ARG, and ARG could be released to the environment even though the wastewater samples have been proceeded in wastewater treatment plants (WWTP). The qPCR assay can be used as the method to monitor the AMR status in a hospital and the spreading potency to the environment using the HWW.