A convenient colorimetric assay for the quantification of porcine epidemic diarrhea virus and neutralizing antibodies
A convenient colorimetric assay for the quantification of porcine epidemic diarrhea virus and neutralizing antibodies
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DOI：
10.1016/j.jviromet.2018.09.003
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发表时间：
2018-12-01
影响因子：
3.1
通讯作者：
Ramamoorthy, Sheela
中科院分区：
文献类型：
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作者：
Singh, Pankaj;Singh, Gagandeep;Ramamoorthy, Sheela
由猪流行性腹泻病毒（PEDV）引起的新生儿肠炎是造成养猪业高死亡率和经济损失的重要原因。诊断实验室通常要求进行病毒中和（V/N）试验，以评估保护性抗体。然而，由操作员对病毒致细胞病变效应进行目视评估以确定抗体滴度或用于病毒定量是一个繁琐、主观和耗时的过程，特别是当涉及高容量检测时。为了提高检测的简易性，在本研究中使用（3-（4，5-二甲基噻唑-2-基）Tr-2，5-二苯基四唑溴化盐）（MTT）（一种测量细胞活力的比色剂）开发并验证了比色病毒中和和TCID 50测定法。将相应的常规测定用作金标准。OD临界值为
Neonatal enteritis caused by the porcine epidemic diarrhea virus (PEDV) is an important cause of high mortality and economic losses to the swine industry. Virus neutralization (V/N) assays are commonly requested in diagnostic laboratories for the assessment of protective antibodies. However, the visual assessment of viral cytopathic effects by operators to determine antibody titers or for viral quantification is a tedious, subjective and time-consuming process, especially when high volume testing is involved. To improve the ease of testing, a colorimetric virus neutralization and TCID50 assays were developed and validated in this study using (3-(4,5-dimethylthiazol-2-yl) Tr-2,5-diphenyltetrazolium-bromide) (MTT), a colorimetric agent which measures cell viability. The respective conventional assays were used as the gold standards. An OD cut off value of