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酶联免疫吸附测定法在绵羊恶性腐蹄病血清学诊断中的应用

Application of ELISA to the serological diagnosis of virulent ovine footrot.

作者信息

Whittington R J, Egerton J R

机构信息

Elizabeth Macarthur Agricultural Institute, NSW Agriculture, Menangle, Australia.

出版信息

Vet Microbiol. 1994 Jul;41(1-2):147-61. doi: 10.1016/0378-1135(94)90144-9.

Abstract

The sensitivity and specificity of an ELISA that detects antibody in the serum of sheep against Dichelobacter nodosus was investigated. The sensitivity of the assay was determined in two trials in which sheep free of virulent footrot were exposed to a virulent strain of D. nodosus. In the first trial up to 96.6% of sheep that developed severe foot lesions were seropositive 2 weeks later. The proportion of seropositive sheep depended on the interval between exposure to D. nodosus and collection of blood samples and reached a maximum 6 weeks after exposure in the first trial and plateaued from 8 weeks after exposure in the second trial. Of sheep with severe lesions present for only 1 week, 40.0% became seropositive, while 98.3% of sheep with severe lesions present for 3 or more weeks became seropositive in Trial 1. Of the unexposed control sheep from the same flock, 93.3% remained seronegative. At least 75% of sheep that were vaccinated with commercial footrot vaccines remained seropositive for at least 24 months after last vaccination and 58.5% remained seropositive for at least 36 months. The specificity of the assay in flocks of sheep from different geographical regions was determined also. Overall, 95.2% of 2978 sheep free of virulent footrot were seronegative. However the rate of false positive serological reactions ranged from 1.6% to 11.3% between flocks and was higher in older sheep (7.0%) compared with sheep less than 1.5 years old (2.7%). Passively acquired colostral antibodies against D. nodosus were measured in 22 lambs and did not persist beyond 55 days mean age. Non-specific reactions with cross reactive antigens were proposed to explain age related changes in serological results. Applications of this ELISA in footrot control programmes are discussed.

摘要

对一种用于检测绵羊血清中抗结节拟杆菌抗体的酶联免疫吸附测定(ELISA)的敏感性和特异性进行了研究。在两项试验中测定了该检测方法的敏感性,试验中使无强毒型腐蹄病的绵羊接触强毒型结节拟杆菌菌株。在第一项试验中,出现严重蹄部病变的绵羊中有高达96.6%在2周后血清呈阳性。血清阳性绵羊的比例取决于接触结节拟杆菌与采集血样之间的间隔时间,在第一项试验中接触后6周达到最高值,在第二项试验中接触后8周达到平稳状态。在仅出现严重病变1周的绵羊中,40.0%血清呈阳性,而在试验1中,出现严重病变3周或更长时间的绵羊中有98.3%血清呈阳性。来自同一羊群的未接触对照绵羊中,93.3%仍为血清阴性。至少75%接种了商用腐蹄病疫苗的绵羊在最后一次接种后至少24个月血清仍呈阳性,58.5%在至少36个月内血清仍呈阳性。还测定了该检测方法在不同地理区域绵羊群中的特异性。总体而言,2978只无强毒型腐蹄病的绵羊中有95.2%血清呈阴性。然而,不同羊群之间假阳性血清学反应率在1.6%至11.3%之间,老年绵羊(7.0%)的假阳性率高于1.5岁以下绵羊(2.7%)。在22只羔羊中检测了被动获得的抗结节拟杆菌初乳抗体,这些抗体在平均年龄55天之后不再持续存在。有人提出与交叉反应抗原的非特异性反应可以解释血清学结果中与年龄相关的变化。讨论了这种ELISA在腐蹄病防控计划中的应用。

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