Quantification of Bacillus Calmette-Guérin (BCG) Tokyo Type I and II Strains in Commercial Lots using Multiplex Real-time PCR

Wichit Thaveekarn, Sunutcha Suntrarachun, Suchitra Khunsap, Jureeporn Noiphrom

Abstract


Japanese bacillus Calmette-Guérin (BCG) strain has been used as a parent strain since 1988 by the Queen Saovabha Memorial Institute, Thai Red Cross Society, to manufacture BCG vaccine in Thailand. In general, the BCG Tokyo strain consists of two BCG subpopulations: type I and type II. The major population is always type I which has a 22-base-pair deletion in the RD16 region. In this study, BCG subpopulations in commercial lots were examined the variation referred to the master seed using modified multiplex real-time PCR in a single tube to quantify two BCG types in commercial lots. Both genotypes were found in almost every lot. The average quantity of type I was 97.26% while that of type II was 2.74%. This method would be useful for quality control of commercial BCG vaccine preparation.

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