腸道病毒基因變異與小兒中樞感染的實(shí)驗(yàn)和臨床研究.pdf

1、Objective: The present study was designed to explore a effective method for diagnosing and genic typing enterovirus ;and to investigate possible associations with VP1 sequence variation and central nerve system infec

2、tion in children. Method: By reverse transcription polymerase chain reaction(RT-PCR), we made use of universal primer to detect enterovirus of central nervous system infection; made use of typing primer to amplify and

3、 sequence the partial VP1 of some EV and then genic typed them according to contrasting GENBANK, furthermore, analyzing the sequence by BLAST software on NCBI; to detect EV71 in clinical specimens by specific primer.

4、 Result: EV was detected positive in 47 specimens from the 63 acute stage children with aseptic infection of central nerve system, the positive rate was 74.6％, it is still the most main pathogen in infection of centr

5、al nerve system. We could amplify 62.0％ positive specimens, random taked 7 products to clone and sequence ,then typed them according to contrasting GENBANK successfully, homology was more than 97.0％. We found 1％～3％ v

6、ariation compared the VP1 sequence with GENBANK, and four definite VP1 sequence of COXB3 mutated on 856th amino acids site. 2 specimens caused by EV71 were detected in Central nervous system infection and 1 specimen ca

7、used by EV71 was detected in hand-foot-mouth disease. Conclusion: ① We could make use of universal primer to detect EV of central nervous system infection by reverse transcription polymerase chainreaction(RT-PCR), the m

8、ethod is specific, sensitive and time-saving. ② The method of typing EV by amplifying VP1 partial sequence and according to contrasting prototype in GENBANK is significant.③The mutation of amino acids on 856th amino ac

9、ids site from glutamine to histidine improve possbly the pathogenicity of COXB3, we presume this mutation cause the virulence and affinity to host acceptor changed ,furthermore ,lead to severe damage of central nervous

10、system, but degree of damage which the same serotype caused is different , and exactd to be further searched.④Central nervous system infection and hand-foot-mouth disease mechanism neecaused by EV71 diffused in Qingdao