Abstract

DETECTION OF BREVUNDIMONAS DIMINUTA BY POLYMERASE CHAIN REACTION IN MEMBRANE FILTER VALIDATION

Bhushan.B*, Uday.D, Mohan.C, Abhay.H, Kakasaheb.M

Abstract

The aim of present work was to develop the PCR based microbial testing method for detection of Brevundimonas diminuta (ATCC 19146) for Bacterial challenge test and compare the results of the conventional microbial testing method and the PCR based microbial testing method. It involved the preparation of the sample for the conventional and PCR based microbial testing, microbial testing by conventional method, PCR based microbial testing (extraction of DNA from bacterial cell and PCR thermal cycling, visualization of PCR product by agarose gel electrophoresis and comparison of results of conventional microbial testing method with PCR based microbial testing method. This technical report is intended to provide a systematic approach to selecting and validating the most appropriate filter for a sterilizing filtration application. Early, careful screening of potential filter types and configurations can result in fewer technical and regulatory problems, fewer delays, more efficient product processing and greater sterility assurance.

Keywords: PCR, Brevundimonas diminuta, PCR thermal cycling, agarose gel electrophoresis, sterilizing filtration.