Development of a blocking ELISA for Caprine parainfluenza virus type
文献类型: 外文期刊
第一作者: Mao, Li
作者: Mao, Li;Li, Wenliang;Zhou, Tianci;Yang, Leilei;Hao, Fei;Li, Jizong;Zhang, Wenwen;Jiang, Jieyuan;Mao, Li;Luo, Xuenong
作者机构:
关键词: Caprine parainfluenza virus type 3;Blocking ELISA;Monoclonal antibodies;Hybridoma screening
期刊名称:JOURNAL OF VIROLOGICAL METHODS ( 影响因子:2.014; 五年影响因子:2.001 )
ISSN: 0166-0934
年卷期: 2017 年 250 卷
页码:
收录情况: SCI
摘要: Caprine parainfluenza virus type 3 (CPIV3) is a novel pathogen mainly causing respiratory diseases in goats. At present, there are no high throughput and rapid testing methods available for epidemiological investigation. In this study, we designed a modified method for selection of hybridomas that secrete monoclonal antibodies (mAb) specific for CPIV3. The monoclonal antibodies were obtained by combination of indirect enzyme-linked immunosorbent assay (iELISA) and blocking ELISA (bELISA). The technique was efficient to determine each mAb with specificity and sensitivity. One bELISA was validated for the serological diagnosis of CPIV3. After optimization conditions were established, a total of 205 reference goat sera were tested in parallel by bELISA and by virus neutralization (VN) for their relative performances. The cut-off point was ultimately defined as 33.6% by ROC curve analysis. The bELISA specificity and sensitivity were 99.2% and 98.7%, respectively, and agreement with the VN test was > 99.0%. Furthermore, testing another 2919 goat sera by bELISA demonstrated 39.3% prevalence in the goat population, more sensitive than HI detection. This new bELISA would offer higher throughput, sensitivity, and specific detection for CPIV3, and will be of great value not only for surveillance, but also for monitoring the efficiency of vaccination programs in the future.
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