New sandwich-type enzyme-linked immunosorbent assay for human MxA protein in a whole blood using monoclonal antibodies against GTP-binding domain for recognition of viral infection

使用针对 GTP 结合域的单克隆抗体对全血中人类 MxA 蛋白进行新型夹心型酶联免疫吸附测定来识别病毒感染

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作者:Mizuho Kawamura, Akira Kusano, Akiko Furuya, Nobuo Hanai, Hideki Tanigaki, Akihito Tomita, Akira Horiguchi, Kyosuke Nagata, Toshiko Itazawa, Yuichi Adachi, Yoshie Okabe, Toshio Miyawaki, Hiroaki Kohno

Conclusion

We have developed a new ELISA for measuring MxA protein in human whole blood using mAbs specific for the GTP-binding domain of MxA. This ELISA has analytical performance enough for routine clinical assay and can be used in detecting the possibility of viral infection.

Methods

A sandwich ELISA suitable for the measurement of human MxA protein in whole blood was developed using mouse monoclonal antibodies (mAbs) raised against the GTP-binding domain of human MxA protein. Prior to the assay, the whole blood sample was treated with special buffer to extract the MxA protein, improve its stability, and avoid interference from hemoglobin.

Results

This ELISA meets all the requirements for use in routine clinical assays, especially in terms of sensitivity (detection limit: 1.3 ng/ml whole blood), accuracy (recovery: 93.0-100.0%), and rapidity (<1.5 h). The present ELISA had a sensitivity of 100% and a specificity of 100% for viral infection when compared to samples from healthy control and 87.1% and 90.9% when compared to samples from the bacterial infection group.

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