检测水禽源细小病毒的胶体金试纸条的研制

发布时间：2018-03-26 05:07

本文选题：鹅细小病毒　切入点：番鸭细小病毒　出处：《中国预防兽医学报》2017年09期

【摘要】：为建立水禽源细小病毒的快速检测方法,本研究采用纯化的小鹅瘟病毒(GPV)免疫BALB/c小鼠,取免疫小鼠脾细胞与SP2/0骨髓瘤细胞进行融合,筛选到21株能够稳定分泌抗GPV的单克隆抗体(MAb),将1株MAb纯化后与胶体金结合,喷涂金标垫,与另一株纯化的MAb配对,制成检测水禽源细小病毒的胶体金试纸条。特异性试验表明,鹅细小病毒、番鸭细小病毒和新型番鸭细小病毒的检测结果均为阳性,猪细小病毒、1型鸭甲肝病毒、坦布苏病毒、产蛋下降综合征病毒、H9N2亚型禽流感病毒、番鸭呼肠孤病毒和新城疫病毒等的检测结果均为阴性。该方法对鹅细小病毒、番鸭细小病毒和新型番鸭细小病毒的最低检出量分别为10~(4.7) ELD50/mL、10~(4.7)ELD50/mL和10~(5.7) ELD50/mL。重复性试验良好。对临床粪便样品的检测,该方法比PCR方法简单快速,但是存在漏检的情况。上述结果表明,本实验建立的方法可用于水禽源细小病毒病的初步快速筛查。
[Abstract]:In order to establish a rapid detection method for waterfowl parvovirus, BALB/c mice were immunized with purified Gosling plague virus (GPVV). The spleen cells of the immunized mice were fused with SP2/0 myeloma cells. We screened out 21 McAb which can secrete GPV stably, then purified one MAb, then combined with colloidal gold, sprayed gold standard pad, paired with another purified MAb. A colloidal gold test strip was made for the detection of waterfowl parvovirus. The specificity test showed that goose parvovirus, muscovy duck parvovirus and new muscovy duck parvovirus were positive, porcine parvovirus type 1 duck hepatitis A virus and tambusuvirus were positive. The detection results of egg drop syndrome virus H9N2 subtype avian influenza virus, muscovy duck reovirus and Newcastle disease virus were negative. The minimum detectable quantities of muscovy duck parvovirus and new muscovy duck parvovirus were 10 ~ 4. 7) ELD50 / mL10 / mL and 10 ~ 5. 7% ELD50 / mL respectively. The reproducibility of the method was good. The detection of clinical fecal samples by this method was simpler and faster than that of PCR method, but there were some cases of missed detection. The method established in this study can be used for the preliminary rapid screening of waterfowl parvovirus disease.
【作者单位】：福建省农业科学院畜牧兽医研究所福建省畜禽疫病防治工程技术研究中心/福建省禽病防治重点实验室;厦门市波生生物技术有限公司;
【基金】：现代农业产业技术体系建设专项资金(CARS-42) 公益性行业(农业)科研专项经费(201003012)
【分类号】：S852.65

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