Jianhua Zhang

Shaoxing University, Shao-hsing, Zhejiang Sheng, China

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Publications (3)5.12 Total impact

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    ABSTRACT: Human enterovirus 71 (EV-71) is the main etiologic agent of hand, foot and mouth disease (HFMD). We sought to identify EV-71 specific antigens and develop serologic assays for acute-phase EV-71 infection. A series of truncated proteins within the N-terminal 100 amino acids (aa) of EV-71 VP1 was expressed in Escherichia coli. Western blot (WB) analysis showed that positions around 11-21 aa contain EV-71-specific antigenic sites, whereas positions 1-5 and 51-100 contain epitopes shared with human coxsackievirus A16 (CV-A16) and human echovirus 6 (E-6). The N-terminal truncated protein of VP1, VP16-43, exhibited good stability and was recognized by anti-EV-71 specific rabbit sera. Alignment analysis showed that VP16-43 is highly conserved among EV-71 strains from different genotypes but was heterologous among other enteroviruses. When the GST-VP16-43 fusion protein was incorporated as antibody-capture agent in a WB assay and an ELISA for detecting anti-EV-71 IgM in human sera, sensitivities of 91.7% and 77.8% were achieved, respectively, with 100% specificity for both. The characterized EV-71 VP1 protein truncated to positions 6-43 aa has potential as an antigen for detection of anti-EV-71 IgM for early diagnosis of EV-71 infection in a WB format.
    Virus research. 06/2014;
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    ABSTRACT: Hand, foot, and mouth disease (HFMD) is caused mainly by enterovirus 71 (EV71) and other enteroviruses (EVs) such as Coxsackie A16 in China. EV71 infection can lead to severe clinical manifestations and even death. Other EVs, however, generally cause mild symptoms. Thus, early and accurate distinction of EV71 from other EVs for HFMD will offer significant benefits. A one-step, single tube, duplex RT-PCR assay is described in the present study to detect simultaneously EV71 and other EVs. The primers used for the duplex RT-PCR underwent screening and optimization. The detection threshold was 0.001 TCID(50) /ml for EV71 and 0.01 TCID(50) /ml for other EVs. The positive rate of enterovirus detection in 165 clinical samples reached 68.5%, including 46.1% for EV71 and 22.4% for other EVs. Of all the severe HFMD cases, EV71 was responsible for 85.3% cases. The positive rate of EV71 fell markedly by day 8 after onset. In addition, sequencing of EV71 specific amplicons from duplex RT-PCR revealed that C4a was the predominant subgenotype of EV71 circulating in Nanjing, China. The accuracy and reliability of the assay suggest strongly that the one-step, single tube, duplex RT-PCR will be useful for early diagnosis and monitoring of EV71 and other EV infections. J. Med. Virol. 84:1803-1808, 2012. © 2012 Wiley Periodicals, Inc.
    Journal of Medical Virology 11/2012; 84(11):1803-8. · 2.37 Impact Factor
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    ABSTRACT: The complete VP1 protein of enterovirus 71 (EV71) and a series of truncations were expressed in Escherichia coli and their antigenic characteristics were studied. Immunoblot analysis showed the major immunoreactive region of the VP1 protein was located in the N-terminal portion at position of amino acid (aa) 1-100. The complete VP1 possessed strong cross-reactivity with antisera against coxsackievirus A16 (CA16) and echovirus 6 (Echo6), while the truncated fragment at position 1-100 aa only had weak cross-reactivity. Moreover, an EV71-specific linear epitope at position 94-105 aa was identified using two EV71-specific mAbs (2B9 and 5B7) with indirect ELISA, but could not be recognized by antibodies against EV71 virus particles. The complete and all of truncated VP1 proteins except His-VP1(202-297) and GST-VP1(202-248) failed to elicit a significant neutralizing antibody response in mice. His-VP1(202-297) and GST-VP1(202-248) containing neutralizing epitope(s) could be recognized only by anti-EV71 mouse sera but not rabbit or human sera. These findings may contribute to a further understanding of antigenic characteristics of the capsid protein VP1 and may be helpful to the development of diagnostic reagents and vaccines.
    Virus Research 05/2012; 167(2):337-42. · 2.75 Impact Factor

Publication Stats

14 Citations
5.12 Total Impact Points

Institutions

  • 2014
    • Shaoxing University
      Shao-hsing, Zhejiang Sheng, China
  • 2012
    • Southeast University (China)
      Nan-ching-hsü, Jiangxi Sheng, China