The Molecular Classification And Laboratory Diagnosis Characteristics Of HFMD Enterovirus 71 Type

Hand-foot-and-mouth disease (Hand-foot-mouth disease HFMD), causes the common infections disease by a number of intestinal virus, mainly in infants. Among them with EV71 and the most common. CoxA16 type. In recent years incidence is increasing trends, from beginning March 2008 in China's anhui province fuyang city popular hand-foot-and-mouth disease pathogen, the China centers for disease control and prevention, laboratory examination results confirm that it was caused by enterovirus71 type (EV - 71). Ended on May 7,2008, the number who infected with hand, foot and mouth has reached 16,778,and 28 death. Given the lack of effective hand-foot-and-mouth disease diagnosis laboratory testing is not typical and clinical manifestation, appear easily moved diagnosis, misdiagnosed, delay treatment, concurrent aseptic meningitis, encephalitis, acute atony sexual paralysis, respiratory infections and myocarditis, individual severe illness progress fast, children vulnerable to death. Especially not typical cases of early diagnosis, has a lot of difficulties. In order to understand the jinan area HFMD enterovirus71 type molecular parting, the more effective laboratory testing of diagnostic indices, the HFMD enterovirus71 type molecular classification and characteristics of research laboratory diagnosis, in order to satisfy the need of clinical diagnosis and reduce the morbidity and mortality of children.This project extracted the HFMD patients excrement and cerebrospinal fluid, the application of viral RNA retrovirus polymerase chain reaction and polymerase chain reaction technical testing virus, and carries on the analysis to study the VP1 fragments genotyped and molecular evolution, and studied in laboratory diagnostic feature. At present already obtained results in the paper are as follows:1, Uses the health ministry recommended primer sequence, with Beijing TIANGEN RT-PCR kit available for amplification, with no clear purpose of the mixed zone. It can be used for outbreak period screening is feasible.2, Our laboratory detected nine samples (from jinan, yucheng respectively, pingyin, hebei qinghe, liangshan, XiaJin and gaotang, linqing,2366-LeLing) in nucleotide 2592 area, its homology 98.7-99.1%, as only one patient deeloped the lack of nucleotides.3, Hand-foot-and-mouth patients with and uncomplicated encephalitis of nucleotides 2366-2592 (VP3-VP1 area) without any different4, With health recommended primer EV71 with our self-designed P2, P4 primer sequencing result is consistent, confirmed the jinan district popular with virus strains EV71 virus strains of fuyang area is homologous new design, but this study of two primer amplification 618bp and fragments of 569bp respectively, far longer than health recommended primer amplification 226bp length, for studying the virus's molecular classification and diagnosis has obvious advantages.5, measured the patient with anhui fuyang, all EV71 evolutionary tree in the same plant, the molecular parting is C4 subtype.6, HFMD detection, most children laboratory performance for the total T cells reduce apparent shrinkage, CD4 cell, severe cases, meningitis, encephalitis and brainstem encephalitis CD4 reduced significantly more patients.This research shows that: in the disease from the early children feces and cerebrospinal fluid extraction RNA for RT - PCR detection methods, HFMD patients for children of enterovirus71 etiology detecting its high sensitivity and specificity is strong, have higher value of clinical application. Promotion, help hand-foot-and-mouth disease early diagnosis, reducing misdiagnosis, mortality and morbidity, improve life quality, improve children population quality, reduce medical costs, reduce the economic burden. In the etiology of hand-foot-and-mouth disease detection at the same time, observe the pathogen popular distribution for prevention and control, and provide basis for developing disease, can have a very important role, therefore has the good social efficiency and the profound social significance.This study medicinal and health technology by shandong institute of information retrieval center online retrieval, and absent or mutation involving EV71 virus literature, but did not see the reports are 2759 bits A subtype of C4 missing and nucleotides in 2659, 2695 2530,2884,2983,2994,3073,3103 3040, and 3182 etc,3133,13 loci variable study reports; Have HFMD patients with complicated brainstem encephalitis, pulmonary edema CD4 + T lymphocytes declining reports in the literature, but didn't see a concurrent encephalitis, meningitis patients CD4 lymphocyte assays reported; Nor has he seen have HFMD patients with severe encephalitis complicated with the encephalitis HFMD reported no difference sequence of nucleotides.