| Baclground:Rash and fever syndrome(RFS)is a series of diseases characterized by fever(?37.5? lasting more than one day)and rash(systemic/local skin or mucous membrane)as the main clinical manifestations.The pathogens causing RFS include viruses and bacteria,which mainly are viruses including enterovirus(EV),measles virus(MV),rubella virus(RV),varicella zoster virus(VZV),human parvovirus B19(HPV B19)and dengue virus(DENV).Among them,in addition to MV,RV,human parvovirus B19 is one of the common human pathogens which can lead to a series of diseases,such as infectious erythema,arthritis,temporary aplastic anemia,abnormal pregnancy,and so on.Some reports indicated that HPV B19 infection rate is high among the population,and can caume an outbreak.So the threat to public health by HPV B19 can not be ignored.The laboratory monitoring technology platform of RFS have been established in some provinces and cities,the results have provided the certain background information for the clarification of pathogen spectrum composition and dynamic changes of RFS.But the researches about etiology and diseases of RFS is not systematic,the monitoring quality and sensitivity remain to be improved.In particular,the studies about epidemiology and genetic characteristics of HPV B19 are very limited,and we lack of scientific,systematic,detailed reports on it.Objective:For the first time,this study analyzed the surveillance data of RFS in Shanxi province from 2009 to 2015,and revealed the composition and epidemiological characteristics of the six viral pathogens(EV,MV,RV,VZV,HPV B19 and DENV).At the same time,We conducted the HPV B19 genotyping and genome sequencing by Nest-PCR,analyzed the genetic characteristics and mutations of B19 strain in Shanxi Province,and provided important scientific data for establishing a sensitive HPV B19 nucleic acid detection method,clarifying the disease burden,preventing and controlling HPV B19 related diseases.Methods:1.Data Source:Information about monitoring cases and clinical specimens of RFSin Shanxi Province from 2009 to 2015 were obtained from the Infectious Disease Surveillance Technology Platform Information Management System.2.Epidemiological Analysis:Excel 2013 and SPSS 22 software was used to conduct epidemiological description and analyze the viral pathogenic spectrum and characteristics of RFS.3.Specimen Source:The clinical specimens of HPV B19 infected cases were collected by Shanxi Center for Disease Control and Prevention and Taiyuan Infectious Diseases Hospital.4.Gene Sequencing:The nucleic acid was extracted,and the HPV B19 gene was amplified and sequenced by Nest-PCR.5.Bioinformation Analysis:Sequencing and analysis of genetic characteristics were conducted by Sequencher,Mega and Bioedit software.Results:1.From January 1,2009 to December 31,2015,850 suspected cases and 1114 specimens of RFS infected by the six virus were collected by two sentinel units,Shanxi center for disease control and prevention(CDC)and Taiyuan infectious diseases hospital.The positive rates of the cases and specimens were 59.57%(504/846)and 58.88%(653/1109),respectively.2.The constituent ratio of pathogens causing RFS,in order,were measles virus 192(38.10%),enterovirus 93(18.45%),rubella virus 87(17.26%),varicella zoster virus 83(16.47%),human parvovirus B19 48(9.52%)and dengue virus 1(0.20%).3.The most cases were children and students under the age of 15.There was no significant difference in detection rate of the six virus among male and female,but the pathogen spectrums were slightly different in different age groups.RFS cases showed a seasonal change,most positive cases occurred from March to August.4.In this study,the Nest-PCR was used to detect the specimens of HPV B19 infection in Shanxi province.The results showed that four specimens were positive and the positive rate was 5.97%.By gene sequencing and homologous analyzing,four strains were all belonged to HPV B19 subtype 1a.5.We obtained a nearly complete genome sequence of B19 virus(SX-T1)in Shanxi with the length of 4983bp.No insertion or deletion of nucleotide or amino acid was found.The nucleotide homology between SX-T1 and reference strains was 86.7%?99.3%in the coding region.Among them,the nucleotide homology between SX-T1 and genotype 1 strains was the highest,followed by genotype 2 and genotype 3.In particular,the nucleotide sequence homology between SX-T1 and Chinese HPV B19 strain HZ1 was the highest,up to 99.3%.There were 1,3,0,1 and 1 amino acids mutated in the coding regions of NS,VP1,X,VP2 and 11KD protein,respectively.The nucleotide homology was 98.3%compared with the subtype la prototype strain Wi,and 6,7,2,2 and 3 amino acids were mutated in the five coding regions respectively.Conclusions:1.The viral pathogens causing RFS were mainly measles virus,enterovirus and rubella virus.The most cases were children and students under the age of 15.There was no significant difference in detection rate of virus among male and female,but the pathogen spectrums were slightly different in different age groups.RFS cases showed a seasonal change,most positive cases occurred in March to August.2.In this study,the Nest-PCR was conducted in HPV B19 genotyping and genome sequencing,which were collected in RFS specimens in Shanxi province.The laboratory testing level was improved and the technical support for establishing a sensitive B19 viras nucleic acid detection method was provided.3.By gene sequencing and homologous analyzing,four strains of Shanxi RFS cases were all belong to HPV B19 subtype 1a.We obtained a nearly complete genome sequence of B19 virus(SX-T1),analyzed the genetic characteristics and mutations of B19 strain in Shanxi Province,and provided important scientific data for clarifying the disease burden,preventing and controlling HPV B19 related diseases. |
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