摘要
目的 比较不同品牌的胎牛血清用于HL-60细胞分化,以及配制成调理缓冲液后对肺炎链球菌调理吞噬杀菌力的影响。方法 采用5个不同品牌(品牌1~5)的胎牛血清用于HL-60细胞分化培养液和调理缓冲液的配制,计算分化后细胞活力,并进行肺炎链球菌调理吞噬杀菌试验,比较对照孔菌数和质控血清的调理指数。结果 与品牌1胎牛血清分化细胞的活力相比较,品牌5分化的细胞活力差异具有统计学意义(P=0.023,P<0.05),而品牌2、3、4分化细胞的活力差异均不具有统计学意义(P=0.129、0.186、0.440,P>0.05);使用品牌5分化的细胞,对3、6A、7F、9V、14、18C、19A、19F血清群/型调理指数因为调理吞噬后菌落数过少,无法计算。使用品牌5配制的调理缓冲液,对3、9V、19F血清群/型调理指数因为调理吞噬后菌落数过少,无法计算。结论 为保障肺炎链球菌调理吞噬试验的稳定性,建议对试验过程中使用的胎牛血清进行筛选。
Objective To compare the effects on opsonophagocytic killing against Streptococcus pneumoniae from different brands of fetal bovine serum(FBS) used for HL-60 cells differentiation and in preparation of an opsonization buffer. Methods Different brands of FBS were used for the preparation of HL-60 cell differentiation culture and an opsonization buffer. After the differentiation, the cell viability was calculated, and opsonophagocytic killing against Streptococcus pneumoniae were conducted. The opsonic index was compared with the quality control serums. Results Compared with the cell viability of differentiated cells by using brand 1, there was a stastical difference(P=0.023, P<0.05)by using brand 5, and no statistical differences by using the other brands including 2, 3, 4(P values were 0.129, 0.186, 0.440 separately, P>0.05). The opsonic index of Serotype 3, 6 A, 7 F, 9 V, 14, 18 C, 19 A, and 19 F could not be calculated due to the small amoumt of colonies after opsonophagocytic killing by using brand 5 to differentiate cells. Serotype 3, 9 V, and 19 F could not be calculated due to the small amoumt colonies when brand 5 was used in preparation of an opsonization buffer. Conclusion It is necessary to screen the FBS used in the experiment, in order to guarantee the stability for opsonophagocytic killing assay against Streptococcus pneumoniae.
作者
黄洋
徐潇
李康
李江姣
杜慧竟
郑锐
孙文媛
叶强
HUANG Yang;XU Xiao;LI Kang;LI Jiang-jiao;DU Hui-jing;ZHENG Rui;SUN Wen-yuan;YE Qiang(Division of Respiratory Bacterial Vaccine National Institutes for Food and Drug Control,Key Laboratory of Method and Standardization for Quality Control of Biotechnical Products,Ministry of Public Health, National Center for Medical Culture Collections, Beijing102629, China)
出处
《微生物学免疫学进展》
2019年第5期22-26,共5页
Progress In Microbiology and Immunology
基金
国家重大新药创制项目(2018ZX09101001,2018ZX09738006-006)
“艾滋病和病毒性肝炎等重大传染病防治”科技重大专项(2018ZX10734404)
关键词
肺炎链球菌
调理吞噬
胎牛血清
Streptococcus pneumoniae
Opsonophagocytic assay
Fetal bovine serum(FBS)
