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Objective: To investigate the monitoring points in Hebei popular pre-epidemic period of healthy children with hand, foot and mouth disease (HFMD) in children pathogenic carrying status and major pathogen characteristics analysis, to provide a scientific basis for doing the monitoring and prevention of HFMD. Methods: According to the Hebei Province in 2009 HFMD incidence in the province to establish six municipal monitoring points, including Shijiazhuang, Baoding, Xingtai City, Tangshan, Langfang, Zhangjiakou City. 2 for healthy children and HFMD children sampled survey of the various monitoring points popular pre and popular period. 3 time quantitative PCR method to detect specimens collected genotyping results for the EV-positive specimens for virus isolation on the part of HFMD in children. 4 EV71 and CA16 VP1 region sequencing primers were designed for sequencing and analysis of both the nucleotide and amino acid homology, respectively, at the same time generate phylogenetic tree analysis of pathogenic genes characteristics. 5 application software for data processing and statistical analysis. Results: 1 monitoring points in Hebei Children detection samples collected in this study were collected from 305 healthy children, including the popular pre-157, 148 epidemic period; acquisition and healthy children matched HFMD children with a total of 297 people, including the popular pre-152 epidemic period 142 people. 2 healthy children quantitative PCR genotyping results detected in healthy children 305 people, including 54 EV-positive, the positive rate of 17.70%, mainly healthy children to other EV 83.33% positive children EV71 and CA16 lower positive rate in healthy children, positive children constitute only 5.56% and 11.11%. Healthy children EV infection to other EV EV71 and CA16 only accounts for less proportion. Healthy children at the sampling time is divided into the popular pre-epidemic period, popular early detection of 157 people, 17 EV-positive, the positive detection rate was 10.83%. The epidemic period detect 148 people, 37 EV-positive, the positive rate was 25.00%. Popular pre-epidemic period of healthy children EV infection are other EV based, respectively, accounting for 64.71% and 91.89% of the positive samples, mainly from HFMD pathogen (the main pathogenic refers EV71 and CA16 both combined) in the proportion of two respectively 35.29% and 8.11%, respectively. Epidemic period of healthy children EV positive rate (25.00%) is higher than the popular pre-(10.83%), and the popular pre-epidemic period pathogens constitute their own characteristics, but are other EV. From a regional distribution point of view, EV positive rate descending order of Baoding 28.21%, Zhangjiakou 26.19%, 22.50% in Tangshan, Xingtai 21.43 percent, 14.29 percent in Langfang, Shijiazhuang 8.00%. The popular pre highest positive rate is 40.91% in Zhangjiakou, the highest positive rate of the epidemic period Baoding 61.11%. Xingtai, Baoding, Tangshan epidemic period were significantly higher than the popular pre. Around the city of HFMD pathogens constitute different. Zhangjiakou City, CA16 and other EV considerable number of other municipalities are other EV. From the crowd, the distribution point of view, the gender and age composition of healthy children EV infection and HFMD pathogen has no effect. 3 HFMD children quantitative PCR genotyping results detected children 297, 167 EV-positive, the positive rate of 56.23%, in the sick children, EV71 accounted for 48.50% of the EV-positive number, followed by CA16 and other EV, EV71 CA16 only one person. HFMD children with major pathogen EV71, CA16 and other EV certain proportion, and exist EV71 and CA16 merger to infection. HFMD children with sampling time is divided into the popular pre-epidemic period, epidemic early detection of 152, 79 EV-positive, the positive rate was 51.97%; prevalence of detection of 145 copies, 88 people EV positive, the positive rate was 60.69 %. Comparison on two EV positive rate and HFMD pathogens, the detection rate was no significant difference, but the pathogens constitute a significant difference on two. Constitute all popular pre-CA16 and EV71 share of higher 49.37% and 39.24%, respectively, other EV accounted for only 11.39%, but the epidemic period, EV71 Share constitutes the highest, rising to 56.82%, followed by other EV's share constitute up to 30.68% and of CA16 share dropped to 11.36%. From a regional distribution point of view, the EV-positive test rate is high in the end turn Shijiazhuang 61.36%, 60.98% in Xingtai, Tangshan, 57.50%, 53.49% in Zhangjiakou, Langfang, 53.33%, 45.00% in Baoding. Most popular pre-positive rate is 69.57% Langfang epidemic period the highest positive rate of 90.00% in Tangshan, Tangshan, Xingtai popular period detection rate increased significantly from the previous month, compared to two, around the city of HFMD pathogen composition of different and each with characteristics. Distribution point of view from the crowd, gender and age for HFMD EV detection rate in children and pathogenic constitute no effect. This is consistent with the results of the health of children. 4 healthy children other EV parting detection results for healthy children other EV positive by further other EV parting detected with viral RNA as a template, RT-PCR amplification with the appropriate primers, target fragments location and size based on the primers used and different. A total of 16 cases were detected, which can determine the type seven cases, the sampling dates epidemic period, 2 human CB5 positive, 1 artificially CA6 positive, 4 artificially ECO14 positive. From Baoding, 3 and (2 CB5 positive, 1 artificially CA6 positive) the Xingtai City 3 (all the ECO14 positive), Tangshan 1 (ECO14 positive). 5 virus isolation results of PCR detection for EV-positive children with specimens for virus isolation, 61 were isolated. 36 of EV71 was identified 18 strains of CA16 4 strains of CA10, 2 strains CA4, 1 strain the CA9. 6 sequence analysis to compare 15 the EV71 isolates VP1 Area characteristics, the results show that 94.7% nucleotide homology to 100.0% amino acid homology of 98.7% to 100%; comparison, with A, B genotype VP1 region homology with genotype C close; further comparison of Hebei Province in 2010 are 15 EV71 C4 The subtypes, EV71 consistent with the different provinces of the country and isolated in Hebei Province in 2009, are the the C4 subtypes C4a branch. 13 CA16 isolates nucleotide homology of 90.8% to 98.9% amino acid homology of 99.0% to 100%; international standard strains with genotype A G-10 (U05876) and 2008 in Fuyang, Anhui strain ( EU812514) nucleotide homology and amino acid homology with the high the B1 subtype B genotype representative strains homology, CA16 strain B1 subtypes are divided in two branch . 7 analysis system tree, Hebei Province EV71 strains and C4 subtypes representative strains of the same branch in 2010, one of the the C4a branch including EV71 strains in the past two years, Hebei and China's different regions representative strains; Hebei Province in 2009 and 2010 22 CA16 strains at home and abroad B1 subtype representative strains in the phylogenetic tree belonging to the B1a and B1b two to evolutionary branch, the popular co-evolution. Hebei Province in recent years EV71 and CA16 genotype stability, not change. Conclusion: healthy children EV 12,010 monitoring points carry high rate of epidemic period were significantly higher than the popular pre; around the pathogen constitute different, but mainly to other EV, EV71 and CA16 only account for a smaller proportion, accompanied There are CB5 CA6 ECO14 Hebei Province monitoring point HFMD EV-positive rate in children in the popular pre-epidemic period does not change; pathogens constitute a major EV71 and CA16, other EV accounted for a smaller proportion, but popular pre-epidemic period constitute slightly different, EV71 gradually occupy a major position. Regardless of the health of children or HFMD in children, not gender and age were found to exist on the of EV detection rate and pathogens constitute. 4 EV71 strains in Hebei province in 2010 the C4 subtypes the C4a branch; CA16 for B1 gene isoforms, respectively, in the B1a and B1b two to evolutionary branch. Hebei Province in recent years EV71 and CA16 genotype stability, not change.
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