Isolation and identification of viruses from patient with hand,foot and mouth disease in Chongqing area in 2009 / 重庆医科大学学报

ABSTRACT

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.