Enzyme immunoassays in which biotinillated beta-lactamase is used for the detection of microbial antigens
Enzyme immunoassays in which biotinillated beta-lactamase is used for the detection of microbial antigens
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使用生物素化 β-内酰胺酶检测微生物抗原的酶免疫分析
DOI：
10.1128/jcm.19.3.356-360.1984
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发表时间：
1984
影响因子：
9.4
通讯作者：
S. Wee
中科院分区：
文献类型：
--
作者：
R. Yolken;S. Wee
酶免疫分析的性能特征在很大程度上取决于用于免疫分析的酶-底物系统。β-内酰胺酶（青霉素酰胺-β-内酰胺水解酶EC 3.5.2.6）提供了许多优点，这些优点可能使它们在免疫测定系统中有用。我们将蜡状芽孢杆菌的β-内酰胺酶与生物素连接，并使用生物素化的酶设计用于检测许多微生物抗原的免疫测定系统。其中使用针对流感嗜血杆菌B型的多核糖醇磷酸抗原的抗体的测定系统能够检测0.4至1.6 ng的该抗原。类似地，其中使用腺病毒和生物素连接的β-内酰胺酶的共同抗原的抗体的测定能够检测肠型腺病毒菌株的1和10 50%之间的组织培养感染剂量。当应用于检测轮状病毒时，使用生物素化β-内酰胺酶的类似系统能够检测标准轮状病毒制剂中的少量抗原。该检测方法也可以在37例轮状病毒胃肠炎患儿的36例粪便标本中检出病毒。用肉眼可以很容易地分辨出阳性和阴性样本，并可以使用标准的办公室复印机永久记录定性结果。β-内酰胺酶有希望用于实际的酶免疫测定系统，特别是在昂贵的比色仪器不可用的情况下。
The performance characteristics of enzyme immunoassays are determined to a great extent by the enzyme-substrate system utilized for the immunoassay. Beta-lactamases (penicillin amido-beta-lactamhydrolase EC 3.5.2.6) offer a number of advantages which might make them useful in immunoassay systems. We linked beta-lactamase from Bacillus cereus with biotin and used the biotinillated enzyme to devise immunoassay systems for the detection of a number of microbial antigens. An assay system in which antibodies to the polyribitol phosphate antigen of Haemophilus influenzae type b were used was capable of detecting between 0.4 and 1.6 ng of that antigen. Similarly, an assay in which antibodies to the common antigens of adenoviruses and biotin-linked beta-lactamase were used was capable of detecting between 1 and 10 50% tissue culture infective doses of a strain of enteric-type adenovirus. When applied to the detection of rotavirus, a similar system in which biotinillated beta-lactamase was used was capable of detecting small amounts of antigen in a standard rotavirus preparation. This assay could also detect virus in 36 of 37 stool specimens from children with rotavirus gastroenteritis. The positive specimens could easily be distinguished from negative ones by the naked eye, and a permanent record of the qualitative results could be obtained by the use of a standard office photocopying machine. Beta-lactamases have promise for use in practical enzyme immunoassay systems, especially in situations in which expensive colorimetric instrumentation is not available.
DOI：
10.1016/s0022-3476(82)80173-x
发表时间：
1982
期刊：
The Journal of pediatrics
影响因子：
--
作者：
Yolken,RH;Lawrence,F;Leister,F;Takiff,HE;Strauss,SE
通讯作者：
Strauss,SE