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荧光抗体试验、免疫过氧化物酶试验和酶联免疫吸附测定法在细胞培养中检测轮状病毒的比较效果

Comparative efficacy of fluorescent antibody test, immunoperoxidase test and enzyme linked immuno sorbent assay in detection of rotavirus in cell culture.

作者信息

Barman N N, Nath A J, Neog B K, Das G

机构信息

Department of Microbiology, College of Veterinary Science, Assam Agricultural University, Khanapara, Guwahati, Assam India.

Pasteur Institute of India, Coonoor, Nilgiris, 643 103 Tamil Nadu India.

出版信息

Virusdisease. 2014;25(2):239-42. doi: 10.1007/s13337-014-0196-x. Epub 2014 Feb 7.

Abstract

Rotavirus is prevalent worldwide and has been established as a leading cause of mortality due to severe diarrhoea in neonates. Isolation of the virus is a gold standard method for confirmation of rotavirus infection in the host. Propagation of rotavirus in cell culture is a challenge as in many instances the virus does not produce detectable cytopathic effect. This study was undertaken to evaluate the efficacy of fluorescent antibody test (FAT), immunoperoxidase test (IPT) and sandwich ELISA (S-ELISA) to detect rotavirus antigen propagated in MA 104 cell line. The intensity of fluorescence and colour development for I-FAT and I-IPT was categorized and the ELISA OD values were analyzed. The overall mean of detection were 5.16 ± 0.47, 5.16 ± 0.54 and 5.66 ± 0.33 for I-FAT, I-IPT and S-ELISA, respectively. Significantly less number of samples were positive in the initial one or two passage, which increased up to 100 % from third/fourth passage onwards. The study concluded that I-FAT, I-IPT and S-ELISA were equally effective in detecting propagated rotavirus in cell line, and the former two tests are suitable for in situ demonstration of the virus while the later could be used to assay antigen in cell culture fluid.

摘要

轮状病毒在全球范围内普遍存在,已被确认为新生儿严重腹泻致死的主要原因。病毒分离是确认宿主中轮状病毒感染的金标准方法。在细胞培养中繁殖轮状病毒是一项挑战,因为在许多情况下该病毒不会产生可检测到的细胞病变效应。本研究旨在评估荧光抗体试验(FAT)、免疫过氧化物酶试验(IPT)和夹心酶联免疫吸附测定(S-ELISA)检测在MA 104细胞系中繁殖的轮状病毒抗原的效果。对I-FAT和I-IPT的荧光强度和显色情况进行分类,并分析ELISA的光密度值。I-FAT、I-IPT和S-ELISA的总体检测平均值分别为5.16±0.47、5.16±0.54和5.66±0.33。在最初的一到两代中呈阳性的样本数量明显较少,从第三代/第四代开始阳性率增加到100%。该研究得出结论,I-FAT、I-IPT和S-ELISA在检测细胞系中繁殖的轮状病毒方面同样有效,前两种试验适用于病毒的原位显示,而后者可用于检测细胞培养液中的抗原。

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