Gweon Eunjeong, Choi Chanwoong, Kim Jaeok, Kim Byungkuk, Kang Hyunkyung, Park Taejun, Ban Sangja, Bae Minseok, Park Sangjin, Jeong Jayoung
Biologics Research Division, National Institute of Food and Drug Safety Evaluation, Cheongju, Korea.
Osong Public Health Res Perspect. 2017 Dec;8(6):389-396. doi: 10.24171/j.phrp.2017.8.6.06. Epub 2017 Dec 31.
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.
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.
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.
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.
为克服当前用于卡介苗（BCG）疫苗活力检测的金标准方法——集落形成单位（CFU）测定法的局限性，我们开发了一种新方法来快速、准确地测定卡介苗疫苗的效力。
基于流式细胞术（FACS）并以荧光素二乙酸酯（FDA）作为最合适的荧光染色试剂，本研究分析了17批经皮给药的卡介苗疫苗和5批皮内给药的卡介苗疫苗。通过流式细胞术测得的活细胞百分比以及在细胞计数器上测定的卡介苗疫苗中生物体总数，用于量化活细胞数量。
经皮和皮内卡介苗疫苗的FACS测定法与CFU测定法的Pearson相关系数分别为0.6962和0.7428，表明相关性较高。FACS测定法的变异系数值小于7%，比CFU测定法低11倍。
本研究有助于评估基于FACS测定卡介苗疫苗中活细胞的新效力测试方法。因此，卡介苗疫苗的质量控制可得到显著改善。
