Identification of Burkholderia cepacia complex by PCR: A simple way

Abstract

USP General Chapter <60> for the detection of Burkholderia cepacia complex (Bcc) members in non-sterile products became official in December 2019. This isolation method requires to confirm the identity of any growth found on Burkholderia cepacia Selective Agar (BCSA) by additional identification tests (refer to the Interpretation section). This article presents a singleplex PCR method to rapidly confirm the membership of any microbial grown on BCSA (& other nutrient medium) in the Bcc group. This method is cost effective as it does not require expensive equipment nor reagents; therefore, it can be easily adopted in the industry. We validated this singleplex PCR Bcc identification method with previously published PCR primers with an expanded panel of 37 clinical and environmental Bcc isolates. The sources and repositories of these Bcc isolates include contaminated health products and medical devices, patients infected with cystic fibrosis, the National Microbiology Laboratory (NML) internal bank and the American type culture collection (ATCC). All 37 isolates that belong to the Bcc have been tested positive using our confirmatory identification method. Twenty-two negative controls including 4 isolates belonging to the genus Burkholderia have been tested negative as expected. Our work indicates this singleplex PCR is an efficient confirmatory method for a Bcc identification, and it can successfully supplement USP chapter <60> in Bcc isolate identification found in pharmaceutical products.