Report of an international collaborative study to evaluate the suitability of multiplex PCR as an identity assay for different sub-strains of BCG vaccine.
Current methods for the identification of BCG vaccine in quality control settings involve acid-fast staining with microscopic examination. However, this method is unable to distinguish the many different sub-strains of BCG, or to differentiate BCG strains from virulent members of the Mycobacterium tuberculosis complex. A multiplex PCR (mPCR) which uses six target regions in mycobacteria has been developed to identify specific sub-strains of BCG. This study reports the findings from an international collaborative study to assess the accuracy, robustness and reproducibility of this mPCR method to differentiate BCG sub-strains. The method was found to fulfil these criteria successfully and was able to distinguish BCG sub-strains in vaccine preparations. The majority of the participants in the study generated the expected PCR product profiles indicating the method is also robust.
Markey
Kevin
K
Bacteriology Division, NIBSC-HPA, Blanche Lane, South Mimms, Potters Bar, Hertfordshire EN6 3QG, UK. kevin.markey@nibsc.hpa.org.uk
Ho
Mei M
MM
Choudhury
Babna
B
Seki
Masaaki
M
Ju
Liu
L
Castello-Branco
Luiz R R
LR
Gairola
Sunil
S
Zhao
Aihua
A
Shibayama
Keigo
K
Andre
Murielle
M
Corbel
Michael J
MJ
D001500:BCG Vaccine / Q000235:genetics*; D015373:Bacterial Typing Techniques / Q000379:methods; D004269:DNA, Bacterial / Q000032:analysis; D007391:International Cooperation; D009163:Mycobacterium bovis / Q000145:classification* / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results
D001500:BCG Vaccine; D004269:DNA, Bacterial