DEMONSTRATION OF THE HUMORAL IMMUNE-RESPONSE OF HORSES TO BABESIA-CABALLI BY WESTERN BLOTTING
DEMONSTRATION OF THE HUMORAL IMMUNE-RESPONSE OF HORSES TO BABESIA-CABALLI BY WESTERN BLOTTING
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DOI：
10.1016/0020-7519(92)90011-9
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发表时间：
1992-08-01
影响因子：
4
通讯作者：
DAEMEN, K
中科院分区：
文献类型：
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作者：
BOSE, R;DAEMEN, K
用十二烷基硫酸钠(十二烷基硫酸钠)缓冲液溶解感染巴贝斯虫或正常马的红细胞，用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳法(SDS-PAGE)和Western blotting进行分析。抗原被允许与实验或现场感染卡巴利球虫的马的血清以及未感染的马的血清发生反应。免疫血清识别的主要巴贝斯虫抗原均有表观摩尔浓度。大小分别为141、112、70、50、48、34和30 kDa。免疫荧光抗体试验(IFAT)和补体结合试验(CFT)假阳性分别为3/100马血清和1/33马血清假阳性。因此，需要进一步鉴定和纯化卡巴氏杆菌抗原，以确定目标抗原，以改进酶免疫检测方法。在这种分析方法出现之前，Western blotting可以为诊断卡巴利支原体感染提供一种特定的工具，特别是在CFT和IFAT结果相互矛盾的情况下。
Babesia caballi-infected or normal equine erythrocytes were solubilized in sodium dodecyl sulfate (SDS) buffer and analyzed by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting. Antigens were allowed to react with sera from horses experimentally or field-infected with B. caballi and with sera from non-infected horses. Major babesial antigens recognized by immune sera had apparent mol. wts of 141, 112, 70, 50, 48, 34, and 30 kDa. The polypeptides at 50 and 48 kDa were recognized earliest and throughout infection, but also weakly by 3/100 equine sera tested negative and 1/33 sera tested false positive by the complement fixation test (CFT) and immunofluorescence antibody test (IFAT). Thus, further characterization and purification of B. caballi antigens are required to identify target antigens for an improved enzyme immuno assay. Until such an assay is available, Western blotting can provide a specific tool for the diagnosis of B. caballi infections, particularly in cases of contradicting CFT and IFAT results.