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Journal of Clinical Microbiology, October 2009, p. 3108-3113, Vol. 47, No. 10
0095-1137/09/$08.00+0     doi:10.1128/JCM.00479-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Monoclonal Antibodies to VP1 Recognize a Broad Range of Enteroviruses {triangledown}

Lynn Yihong Miao,1* Christina Pierce,1 Jennifer Gray-Johnson,1 Jill DeLotell,1 Carl Shaw,1 Nate Chapman,1 Elaine Yeh,2 David Schnurr,2 and Yung T. Huang1

Diagnostic Hybrids, Inc., Athens, Ohio 45701,1 California Department of Public Health, Viral and Rickettsial Disease Laboratory, Richmond, California 948042

Received 6 March 2009/ Returned for modification 13 May 2009/ Accepted 14 August 2009

Enteroviruses (EVs) are common seasonal viruses that are associated with a variety of diseases. High-quality monoclonal antibodies (MAbs) are needed to improve the accuracy of EV diagnosis in clinical laboratories. In the present study, the full-length VP1 genes of poliovirus 1 (Polio 1) and coxsackievirus B3 (Cox B3) were cloned, and the encoded proteins were expressed and used as antigens in an attempt to raise broad-spectrum MAbs to EVs. Two pan-EV MAbs were isolated: one raised against Polio 1 VP1 and the other against Cox B3 VP1. The binding sites of both pan-EV MAbs were mapped to an amino acid sequence within a conserved region in the N terminus of Polio 1 VP1 by peptide and competition enzyme-linked immunosorbent assay. Two additional MAbs, an EV70-specific MAb and an EV71/Cox A16-bispecific MAb, developed against EV70 and 71 VP1 proteins, were pooled with the two pan-EV MAbs (pan-EV MAb mix) and tested for their sensitivity and specificity in the staining of various virus-infected cells. The pan-EV MAb mix detected all 40 prototype EVs tested and showed no cross-reactivity to 18 different non-EV human viruses. Compared with two commercially available EV tests, the pan-EV MAb mix exhibited higher specificity than one test and broader spectrum reactivity than the other. Thus, our study demonstrates that full-length Polio 1 VP1 and Cox B3 VP1 can serve as effective antigens for developing a pan-EV MAb and that the pan-EV MAb mix can be used for the laboratory diagnosis of a wide range of EV infections.

* Corresponding author. Mailing address: R&D Department, Diagnostic Hybrids, Inc., Athens, OH 45701. Phone: (740) 589-3437. Fax: (740) 592-9820. E-mail: miao{at}dhiusa.com

{triangledown} Published ahead of print on 6 August 2009.

Journal of Clinical Microbiology, October 2009, p. 3108-3113, Vol. 47, No. 10
0095-1137/09/$08.00+0     doi:10.1128/JCM.00479-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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