中国比较医学杂志Issue(6)：58-62,5.DOI:10.3969/j.issn.1671.7856.2014.006.013
小鼠诺如病毒间接免疫荧光检测方法的建立
Establishment of an indirect immunofluorescence assay for detection of murine norovirus
摘要
Abstract
Objective To establish an indirect immunofluorescence assay for detection of murine norovirus ( MNV) .Methods Mouse leukaemic monocyte macrophage cell line RAW 264.7 cells were infected with MNV-1 and cultured for 36 hours to collect the virus and uninfected cells , and to make antigen glass slides .BALB/c mice were gavaged with MNV-1 (107 TCID50) and infected sera were collected as positive control .The serum was 1:10 diluted and used for measuring MNV antibody by immunofluorescence assay ( IFA ) .80 serum samples were tested using the two methods , IFA and ELISA, and the discrepant samples were validated by Western blotting .Results RAW264.7 cells were infected with MNV-1 for 36-48 h, showing an infection rate of 60% of the cells, and the cells infected for 36 h were preferred.IFA method was used to detect the serum with MNV-1 infection and showed that the antibody content was gradually increased at one week after infection , reaching a maximum antibody concentration at 4 weeks after infection , and maintained a stable level later .The mouse serum at four weeks after MNV-1infection was used as positive quality control . Among the 80 serum samples , 27 positive and 53 negative cases were detected by IFA method , and 32 positive and 48 negative cases were detected by ELISA .The five discrepant samples were verified by Western blotting , resulted in 3 positive and 2 negative cases . The coincidence rate of IFA was 96.0% and that of ELISA methods was 97.5%. Conclusions Basically, immunofluorescence assay can be used to detect the MNV-1 infection in mice, although false negative result may occur occasionally .IFA and ELISA detection can be selected as initial screening measures , and use Western blot assay to verify the discrepant samples .关键词
间接免疫荧光法/小鼠诺如病毒Key words
Indirect immunofluorescence detection/Murine Norovirus分类
医药卫生引用本文复制引用
田胜男,佟巍,张丽芳,常慧,李雨函,苏静芬,刘先菊,向志光,刘云波..小鼠诺如病毒间接免疫荧光检测方法的建立[J].中国比较医学杂志,2014,(6):58-62,5.基金项目
国家科技支撑计划“实验动物质量保证条件和认可评价关键技术研究与示范”（2011BAI15B03）；国家科技支撑计划“实验动物质量检测关键技术研究”（2013BAK11B01-15）。 （）
