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应用单克隆抗体对抗亨德拉和尼帕病毒的抗原捕获 ELISA 检测。

Use of monoclonal antibodies against Hendra and Nipah viruses in an antigen capture ELISA.

机构信息

Special Pathogens Branch, Division of Viral and Rickettsial Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia, USA.

出版信息

Virol J. 2010 Jun 3;7:115. doi: 10.1186/1743-422X-7-115.

Abstract

BACKGROUND

Outbreaks of Hendra (HeV) and Nipah (NiV) viruses have been reported starting in 1994 and 1998, respectively. Both viruses are capable of causing fatal disease in humans and effecting great economical loss in the livestock industry.

RESULTS

Through screening of hybridomas derived from mice immunized with gamma-irradiated Nipah virus, we identified two secreted antibodies; one reactive with the nucleocapsid (N) protein and the other, the phosphoprotein (P) of henipaviruses. Epitope mapping and protein sequence alignments between NiV and HeV suggest the last 14 amino acids of the carboxyl terminus of the N protein is the target of the anti-N antibody. The anti-P antibody recognizes an epitope in the amino-terminal half of P protein. These monoclonal antibodies were used to develop two antigen capture ELISAs, one for virus detection and the other for differentiation between NiV and HeV. The lower limit of detection of the capture assay with both monoclonal antibodies was 400 pfu. The anti-N antibody was used to successfully detect NiV in a lung tissue suspension from an infected pig.

CONCLUSION

The antigen capture ELISA developed is potentially affordable tool to provide rapid detection and differentiation between the henipaviruses.

摘要

背景

亨德拉病毒(HeV)和尼帕病毒(NiV)分别于 1994 年和 1998 年爆发。这两种病毒都能导致人类致命疾病,并对畜牧业造成巨大经济损失。

结果

通过对用伽马射线辐照的尼帕病毒免疫的小鼠杂交瘤进行筛选,我们鉴定出两种分泌抗体;一种与核衣壳(N)蛋白反应,另一种与亨德拉病毒的磷蛋白(P)反应。尼帕病毒和亨德拉病毒之间的表位作图和蛋白序列比对表明,N 蛋白羧基末端的最后 14 个氨基酸是抗 N 抗体的靶标。抗 P 抗体识别 P 蛋白氨基末端一半的表位。这些单克隆抗体被用于开发两种抗原捕获 ELISA,一种用于病毒检测,另一种用于区分尼帕病毒和亨德拉病毒。两种单克隆抗体的捕获检测的检测下限均为 400 pfu。抗 N 抗体成功地检测到感染猪的肺组织悬浮液中的尼帕病毒。

结论

开发的抗原捕获 ELISA 是一种具有成本效益的快速检测和区分亨德拉病毒的工具。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f608/2896928/085706a697f4/1743-422X-7-115-1.jpg

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