Objective:Toinvestigate the etiological agents of the outbreak ofhand-foot-mouth disease(HFMD)in ChongQing area from April to September in 2009.Methods:100 specimens(including cerebral spinal fluid

vesicle fluid

stool

throat swabs

anal swab) were collected from 47 patients with HFMD during the epidemic season.The specimens were inoculated into RD cells for isolation of enteroviruses(EV)

The supernatant of cytopathic effect(CPE)were detected by RT-PCR with universal primers of enterovirus

genus-specific primers of EV71 and genus-specific primers of Coxsackie A16

and the amplified products were sequenced to determine the outbreak of the pathogen enteroviruses(EV)

The sequencing results will be compared with international representatives BrCr strain to identify the enteroviruses(EV)by Bioedit biological software.Results:In the 100 cell cultures of clinical specimens of children HFMD

apparent Cytopathic were observed in 7.The results of RT-PCR showed that seven specimens were positive with intestinal universal primers

and three were positive results with EV71-specific primers

which were from two cases of out-patient children and one with clinical diagnosis of suspected case

while the negative results were obtained with CA16-specific primers for all the samples.The recovery of the PCR products were cloned and the sequencing identification confirmed as EV71 virus

while sequencing analysis of homology-related in the results.Conclusion:EV71 were the major etiological agents of the HFMD outbreak in Chongqing from February to April in 2009.