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Foot and mouth disease (Hand-foot-mouth disease, HFMD) is a common variety of intestinal infectious disease caused by a virus, infant and child morbidity based. Among the most common type EV71 and CoxA16. The increasing trend of incidence in recent years, from March 2008 began in our Fuyang HFMD epidemic pathogens, the Chinese Center for Disease Control laboratory tests, the results confirmed by enterovirus 71 (EV -71) caused. As of May 7, 2008, the number of patients infected with foot and mouth has reached 16,778 people, killed 28 people. Given the lack of effective HFMD laboratory diagnosis, clinical manifestations becoming atypical prone to misdiagnosis, delayed treatment, complicated by aseptic meningitis, encephalitis, acute flaccid paralysis, respiratory infections and myocarditis, etc., individual children with severe progression fast, easy death. In particular, the early diagnosis of atypical cases, there is a big difficulty. To understand the Jinan region HFMD enterovirus type 71 molecular typing, research more effective laboratory diagnostic index, so the hand, foot and mouth disease enterovirus type 71 molecular typing characteristics and laboratory diagnostic study to meet the needs of clinical diagnosis, reduce child morbidity and mortality. This topic is designed to our hospital HFMD children's faeces and CSF viral RNA extraction methods, reverse transcriptase polymerase chain reaction and polymerase chain reaction to detect viruses, and its VPl fragment analysis to study the genotyping and molecular evolution, and laboratory diagnostic characteristics were studied. Has achieved the following results: 1, using the Ministry of Health recommended primer sequences, with the Beijing-day root RT-PCR amplification kit, available with a clear purpose of non-hybrid bands. Screening for the outbreak period is feasible. 2, I chamber test 9 samples (respectively from Jinan, Yuncheng, Pingyin, Hebei Qinghe, Yangsan, Xiajin, Gaotang, Linqing, Leling) nucleotides 2366-2592 in the region, and its high homology 98.7-99.1%, only one nucleotide deletion patients. 3, foot and mouth disease complicated with non-nucleotide encephalitis region in 2366-2592 was no different (VP3-VP1 part of the region) 4, with the Ministry of Health recommended EV71 primers with our own design P2, P4 sequencing primer The results are consistent, proven popular in Jinan Fuyang EV71 strains and strains of the virus are homologous, but this study newly designed two pairs of primers amplified fragments were 618bp and 569bp, much longer than recommended by the Ministry of Health primers amplified 226bp in length, for the study of viruses and diagnostic molecular typing has obvious advantages. 5, the measured patient EV71, Fuyang, Anhui strains were associated with the same evolutionary tree, molecular typing of C4 subtypes. 6, hand, foot and mouth disease children laboratory testing found that most of the total T cells showed reduced, CD4 cells was significantly reduced, in severe cases, encephalitis, meningitis and brain stem encephalitis patients more pronounced decrease in CD4. The research showed that: early in the disease and cerebrospinal fluid from children with fecal RNA was extracted for RT-PCR detection method, the children HFMD children etiology of enterovirus type 71 was detected with high sensitivity and specificity, there high clinical value. After promotion to help early diagnosis of foot and mouth disease, reduce misdiagnosis and reduce mortality and morbidity, improve the quality of life of children and improve the population quality, reduce medical costs, reduce the financial burden on the patient. In HFMD pathogen detection while observing the distribution of the pathogen prevalence, prevention and control and to provide the basis for development of disease, can play a very important role, which has a good social and far-reaching social significance. This study was approved by Shandong Province Science and Technology Information Research Institute of Pharmaceutical Health retrieval center line retrieval, and abroad involving EV71 virus deleted or mutated reported in the literature, but there is no lack of C4 subtypes A and 2759 nucleotides in 2530,2659,2695 , 2884,2983,2994,3040,3073,3103,3133,3182 etc. 13 Loci research reports; HFMD brainstem encephalitis, pulmonary edema in patients with CD4 T lymphocyte decline reported in the literature, but not saw encephalitis, meningitis patients reported in the literature CD4 lymphocytes detected; HFMD is observed that in patients with non-complicated by severe encephalitis encephalitis nucleotide sequence HFMD patients who reported no difference.
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