Enterobacterial intergenic consensus sequence polymerase chain reaction as a typing method for Burkholderia (Pseudomonas) cepacia.
OBJECTIVE: We developed a rapid polymerase chain reaction (PCR) method, which utilizes primers from an enterobacterial repetitive consensus sequence, to differentiate among strains of Burkholderia cepacia. We then applied this method to isolates from a device-associated pseudoinfection and also compared the discriminative typing potential with that of another PCR-based method. METHODS: A simplified lysis procedure was used to generate DNA substrate for amplification by a PCR method which utilized primer ERIC2 and a reverse complement of ERIC1R. The alternative PCR-based method employed primers which are directed to Bordetella pertussis repetitive sequences. Amplification products were visually assessed after conventional agarose gel electrophoresis and staining. RESULTS: The ERIC-based method appropriately clustered strains for the device-associated pseudo-infection. In addition, there was consistency in the definition of similar or dissimilar strains between the two PCR---based systems. CONCLUSIONS: The ERIC-based PCR typing method offers sufficient discriminatory power for it to be used for epidemiologic purposes.
Cimolai
Nevio
N
Department of Pathology, British Columbia's Children's Hospital, Vancouver, Canada.
Trombley
Colleen
C