Osong Public Health Res Perspect.  2017 Dec;8(6):389-396. 10.24171/j.phrp.2017.8.6.06.

Development of a New Approach to Determine the Potency of Bacille Calmette–Guérin Vaccines Using Flow Cytometry

• Gweon E
• Choi C
• Kim J
• Kim B
• Kang H
• Park T
• Ban S
• Bae M
• Park S
• Jeong J

• KMID: 2402585
• DOI: http://doi.org/10.24171/j.phrp.2017.8.6.06

Abstract

OBJECTIVES
To circumvent the limitations of the current golden standard method, colony-forming unit (CFU) assay, for viability of Bacille Calmette-Guérin (BCG) vaccines, we developed a new method to rapidly and accurately determine the potency of BCG vaccines.
METHODS
Based on flow cytometry (FACS) and fluorescein diacetate (FDA) as the most appropriate fluorescent staining reagent, 17 lots of BCG vaccines for percutaneous administration and 5 lots of BCG vaccines for intradermal administration were analyzed in this study. The percentage of viable cells measured by flow cytometry along with the total number of organisms in BCG vaccines, as determined on a cell counter, was used to quantify the number of viable cells.
RESULTS
Pearson correlation coefficients of FACS and CFU assays for percutaneous and intradermal BCG vaccines were 0.6962 and 0.7428, respectively, indicating a high correlation. The coefficient of variation value of the FACS assay was less than 7%, which was 11 times lower than that of the CFU assay.
CONCLUSION
This study contributes to the evaluation of new potency test method for FACS-based determination of viable cells in BCG vaccines. Accordingly, quality control of BCG vaccines can be significantly improved.

Keyword

BCG vaccine; vaccine potency; flow cytometry

MeSH Terms

Administration, Cutaneous
BCG Vaccine
Cell Count
Flow Cytometry*
Fluorescein
Methods
Mycobacterium bovis
Quality Control
Stem Cells
Vaccine Potency
Vaccines*
BCG Vaccine
Fluorescein
Vaccines