Rapid detection and diagnosis of human enterovirus caused an encephalitis outbreak in Guangdong,China

Objective To rapidly detect and identify the viral encephalitis caused by human enterovirus.Methods Cerebrospinal fluid(CSF)and stool specimens of the viral encephalitis cases were rapidly screened by using real-time quantitative PCR with universal enterovirus primers.The partial VP1 gene fragment from positive specimens was amplified using semi-nested PCR(Sn-PCR),sequenced and aligned using BLAST with the corresponding subtypes in the NCBI GenBank.Results Out of 32 CSF specimens tested,26(81.3%)were enterovirus positive by real-time PCR,and all 7(100.0%)stool specimens tested were enterovirus positive.Twenty-six out of 33 positive samples were successfully amplified for the target VP1 gene fragment by Sn-PCR.Sequencing analysis showed that 13 of the 15 CSF specimens were ECV30,the other two were ECV9;6 of the 7 stool specimens were ECV30,only one was ECV9.Conclusions The pandemic of viral encephalitis was proved to be an enterovirus outbreak in Luoding city,west Guangdong province of China.Real-time PCR and Sn-PCR are very effective ways for rapid detection and identification of the intestinal viral encephalitis.