Establishment of a CODEHOP RT-PCR method for detecting Alphavirus carried by mosquito

Objective To develop a CODEHOP RT-PCR method for detecting Alphavirus carried by mosquito.Methods According to the amino acid sequences of nonstructural protein of Alphavirus, a pair of CODEHOP primers was designed. Based on which a method of one step RT-PCR was established for detecting Alphavirus. The primer binding sites in different species of Alphavirus and the size of the products were analyzed. The sensitivity and specificity of the method were evaluated according to the result of detecting CHIKV JC2012. Results The CODEHOP RT-PCR method could amplify the CHIKV JC2012 specially, the sequence was completely consistent with the published sequence of CHIKV JC2012 and the limit of detection of nucleic acid was 56.7 pg. Meanwhile, all of the 22 Alphavirus could be find the binding sites for the CODEHOP primers, and all the amplifications were 510~550 bp. Conclusion The CODEHOP RT-PCR method is specific and sensitive which could be used to detect mosquito-brone Alphavirus.