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直接在固定的感染细胞单层上进行酶联免疫吸附测定以评估抗水痘带状疱疹病毒药物的应用。

Use of an enzyme-linked immunosorbent assay performed directly on fixed infected cell monolayers for evaluating drugs against varicella-zoster virus.

作者信息

Berkowitz F E, Levin M J

出版信息

Antimicrob Agents Chemother. 1985 Aug;28(2):207-10. doi: 10.1128/AAC.28.2.207.

Abstract

An in situ enzyme-linked immunosorbent assay (ELISA), performed directly on fixed varicella-zoster virus-infected monolayers, was used to quantitate viral antigen, and by color reduction it was used to evaluate the activity of drugs against varicella-zoster virus. Color production in the ELISA (optical density) was directly related to the dose of input virus. Antigen representing 5 to 10 plaques could be detected 3 days after infection. The ELISA was specific and reproducible, as shown by absorption and repeat experiments, respectively. A color-reduction test by ELISA was compared with the conventional plaque-reduction assay for its ability to measure the antiviral activity of acyclovir, bromovinyldeoxyuridine, trifluorothymidine, and vidarabine against four strains of varicella-zoster virus. In all cases but one the 50% inhibitory doses were lower when measured by ELISA than by the plaque-reduction assay. This in situ ELISA color reduction method had the following advantages over the conventional plaque-reduction assay: the endpoint was an objective measurement; there was less well-to-well variation; the assay was sensitive to changes in plaque size as well as plaque number; it was less labor intensive.

摘要

一种直接在固定的水痘带状疱疹病毒感染单层细胞上进行的原位酶联免疫吸附测定(ELISA),用于定量病毒抗原,并通过颜色还原评估药物对水痘带状疱疹病毒的活性。ELISA中的显色(光密度)与输入病毒的剂量直接相关。感染后3天可检测到代表5至10个蚀斑的抗原。分别通过吸收实验和重复实验表明,ELISA具有特异性和可重复性。将ELISA的颜色还原试验与传统的蚀斑减少试验进行比较,以评估阿昔洛韦、溴乙烯脱氧尿苷、三氟胸苷和阿糖腺苷对四株水痘带状疱疹病毒的抗病毒活性。除一例以外,在所有情况下,通过ELISA测量的50%抑制剂量均低于蚀斑减少试验测量的结果。这种原位ELISA颜色还原方法相对于传统的蚀斑减少试验具有以下优点:终点是客观测量;孔间差异较小;该试验对蚀斑大小和蚀斑数量的变化均敏感;劳动强度较小。

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