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猪丹毒杆菌保护性抗原在酶联免疫吸附测定和乳胶凝集试验中的应用。

Use of the protective antigen of Erysipelothrix rhusiopathiae in the enzyme-linked immunosorbent assay and latex agglutination.

作者信息

Sato H, Yamazaki Y, Tsuchiya K, Aoyama T, Akaba N, Suzuki T, Yokoyama A, Saito H, Maehara N

机构信息

Department of Veterinary Microbiology, School of Veterinary Medicine and Animal Sciences, Kitasato University, Aomori, Japan.

出版信息

Zentralbl Veterinarmed B. 1998 Sep;45(7):407-20. doi: 10.1111/j.1439-0450.1998.tb00810.x.

Abstract

To establish a safe and convenient serodiagnostic method for swine erysipelas, a purified protective protein antigen of Erysipelothrix rhusiopathiae, which included a large amount of protective protein (64 kDa protein), was used for enzyme-linked immunosorbent assay (ELISA) and the latex agglutination (LA) test. In the ELISA, the antisera to four different serovars (1a, 2, 5 and 20) of E. rhusiopathiae exhibit a positive reaction, while antisera to other species of bacteria (Listeria monocytogenes, Staphylococcus aureus, Streptococcus suis, Rhodococcus equi and Corynebacterium pseudotuberculosis) exhibit a negative reaction. In the LA test, the antisera to three different serovars (1a, 2 and 5) of E. rhusiopathiae reacted with P64-sensitized latex beads, while the antiserum to serovar 20 (2553 strain) did not. Moreover, the antisera to other species of bacteria (Listeria monocytogenes, Staphylococcus aureus, Streptococcus suis, Rhodococcus equi and Corynebacterium pseudotuberculosis) did not in this test. Comparing the results of the growth agglutination (GA), ELISA and LA tests of 284 swine sera, there was a high degree of correlation among the results. The detection of anti-E. rhusiopathiae antibodies in the GA, ELISA and LA tests were compared using sera from pigs immunized with P64, alkaline extract (AE) and live-cell vaccine (LV). In all three tests, anti-E. rhusiopathiae antibodies could be detected 1 week after immunization. The serum antibody titre as determined by the LA test increased moderately, as did that by the GA test, while that determined by ELISA increased rapidly. These results suggested that ELISA could be used to monitor changes in anti-E. rhusiopathiae antibody titre and the LA test could be used in the screening test for swine erysipelas.

摘要

为建立一种安全便捷的猪丹毒血清学诊断方法,将包含大量保护性蛋白(64 kDa蛋白)的猪红斑丹毒丝菌纯化保护性蛋白抗原用于酶联免疫吸附测定(ELISA)和乳胶凝集(LA)试验。在ELISA中,针对猪红斑丹毒丝菌四种不同血清型(1a、2、5和20)的抗血清呈现阳性反应,而针对其他细菌种类(单核细胞增生李斯特菌、金黄色葡萄球菌、猪链球菌、马红球菌和伪结核棒状杆菌)的抗血清呈现阴性反应。在LA试验中,针对猪红斑丹毒丝菌三种不同血清型(1a、2和5)的抗血清与P64致敏乳胶珠发生反应,而针对血清型20(2553菌株)的抗血清未发生反应。此外,针对其他细菌种类(单核细胞增生李斯特菌、金黄色葡萄球菌、猪链球菌、马红球菌和伪结核棒状杆菌)的抗血清在该试验中也未发生反应。比较284份猪血清的生长凝集(GA)、ELISA和LA试验结果,结果之间存在高度相关性。使用用P64、碱性提取物(AE)和活细胞疫苗(LV)免疫的猪的血清比较GA、ELISA和LA试验中抗猪红斑丹毒丝菌抗体的检测情况。在所有三项试验中,免疫后1周均可检测到抗猪红斑丹毒丝菌抗体。LA试验测定的血清抗体滴度适度升高,GA试验测定的情况也是如此,而ELISA测定的血清抗体滴度迅速升高。这些结果表明,ELISA可用于监测抗猪红斑丹毒丝菌抗体滴度的变化,LA试验可用于猪丹毒的筛查试验。

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