Development of a tetrazolium salt assay for rapid determination of viability of BCG vaccines
Development of a tetrazolium salt assay for rapid determination of viability of BCG vaccines
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DOI：
10.1016/s0264-410x(99)00023-7
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发表时间：
1999-05-14
期刊：
影响因子：
5.5
通讯作者：
Corbel, MJ
中科院分区：
文献类型：
--
作者：
Kairo, SK;Bedwell, J;Corbel, MJ
按照世界卫生组织（WHO）和欧洲药典（EP）的规定对活牛分枝杆菌BCG（BCG）疫苗进行标准化和控制。用于控制BCG疫苗效力的常规活菌计数通过在固体培养基上培养来进行。这种测定方法不仅耗时，而且可能得到可变的结果。已开发并评价了四唑盐试验，作为测定活微生物数量的潜在附加或替代试验。四唑盐3-（4，5-二甲基噻唑-2-基）-2，5-二苯基溴化四唑（MTT）和2，3-双-（2-甲氧基-4-硝基-5-磺基）-（2 H）-四唑-5-甲酰苯胺（XTT）作为替代底物，在该方法中均比常规活菌计数法得到更快速和可重复的结果。XTT显示出比MTT更高的灵敏度，检测下限约为7 × 10（4）菌落形成单位（c.f.u.）ml（-1）。XTT检测已被证明可有效测定由几种BCG疫苗亚株制备的混悬液的活力，涵盖一系列活单位，无需修改。该测定法易于进行，仅需48小时即可估计活细胞含量，而传统方法需要3周。(C)1999由Elsevier Science Ltd.出版。保留所有权利。
Standardisation and control of the live Mycobacterium bovis BCG (BCG) Vaccine is performed as specified by the World Health Organisation (WHO) and the European Pharmacopoeia (EP). The conventional Viable count for control of potency of BCG vaccine is performed by culturing on solid medium. This assay method is not only time consuming but may give variable results. A tetrazolium salt assay has been developed and evaluated as a potential additional, or replacement, test for determining number of viable organisms. The tetrazolium salts 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and 2,3-bis- (2-methoxy-4-nitro-5-sulphenyl)-(2H)-tetrazolium-5-carboxanilide (XTT) used as alternative substrates in the ass ay both gave more rapid and reproducible results than the conventional viable count. XTT showed greater sensitivity than MTT with a lower detection limit of about 7 x 10(4) colony forming units (c.f.u.) ml(-1). The XTT assay has proven effective for determining viability of suspensions prepared from several BCG vaccine substrains, covering a range of viable units, without the need for modification. This assay is easily performed and takes just 48 h to produce an estimate of viable cell content compared with 3 weeks for the conventional method. (C) 1999 Published by Elsevier Science Ltd. All rights reserved.