Yokoyama T, Kimura K, Tagawa Y, Yuasa N
National Institute of Animal Health, Ibaraki, Japan.
Clin Diagn Lab Immunol. 1995 Mar;2(2):172-6. doi: 10.1128/cdli.2.2.172-176.1995.
Antisera were raised in rabbits against three peptides, representing amino acid sequences 150 to 159, 165 to 174, and 213 to 226 of mouse prion (PrP), which were synthesized by using a multiple antigenic peptide (MAP) system. The reactivities of these sera to PrP were examined by an enzyme-linked immunosorbent assay (ELISA), Western immunoblotting (WB), and immunohistochemical procedures. The results of both ELISA and WB showed that antisera to peptide sequence 150 to 159 (Ab150-159) did not react with purified mouse PrP. On the other hand, sera to the sequence 165 to 174 (Ab165-174) reacted weakly with purified mouse PrP, as detected by WB but not by ELISA. However, antiserum to peptide sequence 213 to 226 (Ab213-226) reacted strongly with mouse, Syrian hamster, and sheep PrP by WB and with mouse PrP as shown by the results of ELISA. Moreover, Ab213-226 clearly detected PrP immunohistochemically in mouse, Syrian hamster, and sheep brains affected with scrapie as well as in the brain of a cow with bovine spongiform encephalopathy. From these data, we conclude that rabbit antiserum against the MAP representing amino acid sequence 213 to 226 of mouse PrP is useful as a diagnostic tool for prion disease of animals.
用三种肽对兔子进行免疫，制备抗血清。这三种肽分别代表小鼠朊病毒（PrP）的氨基酸序列150至159、165至174和213至226，它们是使用多抗原肽（MAP）系统合成的。通过酶联免疫吸附测定（ELISA）、蛋白质免疫印迹法（WB）和免疫组织化学方法检测这些血清与PrP的反应性。ELISA和WB的结果均显示，针对肽序列150至159的抗血清（Ab150-159）不与纯化的小鼠PrP发生反应。另一方面，针对序列165至174的血清（Ab165-174）与纯化的小鼠PrP发生弱反应，WB检测到了这种反应，但ELISA未检测到。然而，针对肽序列213至226的抗血清（Ab213-226）通过WB与小鼠、叙利亚仓鼠和绵羊的PrP发生强烈反应，ELISA结果也显示其与小鼠PrP发生反应。此外，Ab213-226在免疫组织化学上能清晰地检测到感染羊瘙痒病的小鼠、叙利亚仓鼠和绵羊大脑以及患牛海绵状脑病的牛大脑中的PrP。根据这些数据，我们得出结论，针对代表小鼠PrP氨基酸序列213至226的MAP的兔抗血清可作为动物朊病毒病的诊断工具。
