Randhawa A S, Stanton G J, Green J A, Baron S
J Clin Microbiol. 1977 May;5(5):535-42. doi: 10.1128/jcm.5.5.535-542.1977.
A liquid antibody microculture plaque assay and the variables that govern its effectiveness are described. The assay is based on the principle that low concentrations of homologous antibody can inhibit secondary plaque formation without inhibiting formation of primary plaques. Thus, clear plaques that followed a linear dose response were produced. The assay was found to be more rapid, less cumbersome, and less expensive than assays using agar overlays and larger tissue culture plates. It was reproducible, quantitative, and had about the same sensitivity as the agar overlay technique in measuring infectious coxsackievirus type B-3. It was more sensitive in assaying adenovirus type 3 and Western equine encephalomyelitis, vesicular stomatitis, Semliki forest, Sendai, Sindbis, and Newcastle disease viruses than were liquid, carboxymethylcellulose, and methylcellulose microculture plaque assays. The variables influencing sensitivity and accuracy, as determined by using coxsackievirus type B-3, were: (i) the inoculum volume of virus; (ii) the incubation period of virus; and (iii) the incubation temperature.
本文描述了一种液体抗体微量培养蚀斑测定法及其影响有效性的变量。该测定法基于这样的原理：低浓度的同源抗体可抑制继发性蚀斑形成，而不抑制原发性蚀斑的形成。因此，产生了呈线性剂量反应的清晰蚀斑。结果发现，该测定法比使用琼脂覆盖层和更大组织培养板的测定法更快、更简便且成本更低。它具有可重复性、定量性，在检测B - 3型传染性柯萨奇病毒时，其灵敏度与琼脂覆盖层技术大致相同。在检测3型腺病毒、西部马脑炎病毒、水疱性口炎病毒、塞姆利基森林病毒、仙台病毒、辛德毕斯病毒和新城疫病毒时，它比液体、羧甲基纤维素和甲基纤维素微量培养蚀斑测定法更灵敏。通过使用B - 3型柯萨奇病毒确定的影响灵敏度和准确性的变量为：(i)病毒接种量；(ii)病毒孵育期；(iii)孵育温度。
