Determination of materials genome in Varicella vaccine in Vietnam by PCR method

Abstract

Background/Purpose: National Institute for Control of Vaccines and Biologicals is
reponsible for its quality control. Identity test is compulsory in quality control tests. WHO
currently recommends Plaque Forming Unit Test for identity test of varicella vaccine. The
limitation is requirement of specific antibody and reference standard of manufacturer
because International Reference vaccine and antibody are not available. At NICVB, the
research has established an identity test protocol¬ by PCR to minimize testing time and
dependence to manufacture’s reagents
Methods: PCR methods, Validation identity methods.
Results: Primers are designed to target stable domain ORF-7, length of product is 280
nucleotide. PCR reaction contains: 12.5 µl master mix, 5.5 µl H2O, 1 µl forward
primer/reverse primer, 5 µl template and annealing temperature is 500C, 30 cycles. The
protocol is optimized, following validation of robustness and limit of detection. Identity
test protocol is applied in vaccine quality control.
Conclusion: The study has designed primers targeting ORF-7 gene for identity test of varicella
virus. The protocol for identity test is validated and meet the criteria of robustness, specificity
and limit of detection.