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Expression of caspid protein VP1 for use as antigen for the diagnosis of enterovirus 71 infection

✍ Scribed by Shin-Ru Shih; Yi-Shuane Li; Chiuan-Chian Chiou; Pin-Chau Suen; Tzou-Yien Lin; Luan-Yin Chang; Yhu-Chering Huang; Kuo-Chien Tsao; Hsiao-Chen Ning; Tzong-Zeng Wu; Err-Cheng Chan

Publisher
John Wiley and Sons
Year
2000
Tongue
English
Weight
916 KB
Volume
61
Category
Article
ISSN
0146-6615

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✦ Synopsis

To produce enterovirus 71 antigen for diagnostic purposes, the gene encoding the entire capsid protein VP1 was amplified by reverse transcription-polymerase chain reaction (RT-PCR), cloned and expressed in Escherichia coli as a polyhistidine fusion protein. Western blotting experiments with sera from patients with enterovirus 71 infection indicated that immunoglobulin G (IgG) and IgM antibodies bound to a single polypeptide VP1. According to these results, IgM anti-VP1 appeared in sera of patients with a symptomatic enterovirus 71 acute infection, whereas IgG anti-VP1 was present in sera of past infection. This finding suggests that detecting IgG and IgM immune responses against linear epitopes of recombinant VP1 is an effective means of determining the different phases of enterovirus 71 infection. In addition, sera containing coxsackie virus 16 (CA16) antibodies did not cross-react with the recombinant VP1 of enterovirus 71, despite the homology between VP1 proteins of both viruses. Comparison with reference PCR and neutralization assays showed these antibody tests to be appropriate for the serodiagnosis of enterovirus 71 infection.

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