| Rabies, a highly lethal meningitis caused by lyssavirus （LYSSA） infection of the cental nerves system, is one of the earliest Epidemic diseases that have ever discovered and recognized by human.Rabies is also the most deadly disease resulted almost in 100% mortality rate, represents as two main clinical types, manic and paralysis.Different pathogenicities of LYSSA determined by different genotypes. Rabies spreading in mammals, as in Dogs and cats, are mainly typl rabies virus, clinically represented as manic. The phenotypes of Rabies spreading in bats are relatively weak, however more complex and diverse, clinically represented as paralysis.LYSSA is an unsegmented 12KB genome length, single-strand negative RNA virus, classified as a member of Rhabdoviridae class. Viral genome from 3’ to 5’ end in turn encoded 5 structural proteins:N, P, M, G, L. Although some new undetermined LYSSA were reported in recent years, only 14 virus out of 7 LYSSA genotypes are so far recognized.The current study constructed the technology of LYSSA Molecular typing technique, rabies surveillance information report, data management system, as well as laboratory rapid virus detection method, fulfilled the needs of establishing a cooperation platform for the information and resources communication and rabies study information networking transferring among multiple departments and different agencies. The current study relied on the "technique", surveillance and analyzing, recovered the population and geographical distribution of rabies virus strains in China. We had successfully discovered the first case of imported laboratory confirmed case of rabies, analyzed its molecular biological characteristics and traced the source by traceability analysis. We had also monitored and got the whole genome sequence of the epidemic strains of the Arctic correlation group. Moreover, we also systematic compared and analyzed the character of different gene type groups in China.This study also provides an important technical support for the further improvement of the network surveillance ability of rabies-related laboratories in China, reveals inherent disciplines, epidemic features and the transmission factors of rabies in China.Charpterl. Establishment of network surveillance technology for rabies laboratoryFirstly, the epidemiological characteristics and distribution characteristics of rabies in China were analyzed. Data shown the overall situation of rabies in China showed a downward trend in 2013. It was the sixth straight year of epidemic declining since the peak of the epidemic period was found in 2007.Most cases of rabies were found in rural areas in China, showing an obvious regional difference. According to the report, most cases were found around in the parts of the south of the Yangtze River. Although the epidemic situation is similar to the recent surveillance, an obvious decreasing of differences is shown compared with the north area. This indicates an obvious rabies epidemic spreading. The rapid spreading and higher incidence of rabies should warn us to make adequate precautions.In order to improve the laboratory surveillance ability of rabies, we used PCR and gene sequencing technology, successfully established rapid identification method of LYSSA detection system of nested RT-PCR, LYSSA whole genome sequencing system, aiming at improve the specificity of different rabies genus detection, as well as laboratory surveillance.In addition, for a further improvement in the ability of Rabies laboratory network surveillance, we also established a multiple departements-sharing epidemiology data base, pathogenic database and gene database.Charpter2. The application of laboratory network surveillance platform for rabiesStudy monitored and analyzed population and geographical distribution of rabies virus strains in China, completed Specimen collection and detection, Sequence determination and homology comparison and phylogenetic analysis, Sequence determination and homology comparison and phylogenetic analysis, completed the collection and detection of specimens, sequence determination and homology comparison and phylogenetic analysis.8441 samples were totally involved, including 384 positive samples, separately from hosts like human, Canine animals, cats, ferretbadger, mouse and cattle. Almost all the N-gene sequence with clear background information were up to now combined with new ones,452 out of which in total were analyzed with phylogenetic analysis. Cases were collected from 26 provinces, lasting 47 years（1969-2015）.Results shown that rabies in China can be devised into 6 virulent strain groups, I-VI. Among which, China I is currently the main epidemic type. Epidemic range covered almost northwest and south west China. According to the trend, the west and northeast parts of china will so only spread and affected.Besides China I, China II is also a wide spread, large numbered strains of population. The current research showed that China II had spread over 14 provinces. Indicating that the number and range were co-increases in recent years. As China I and China II are groups hosting mainly in dogs, strain groups of ferrets widely spread around Zhejiang and Jiangxi are mostly belongs to China II, except a very few, data shown an obvious distribution difference.Numbers of China III is far more less than China I and II, however the range covers more than we have estimated 9 provinces including, Xinjiang and Inner Mongolia cows, sheep and camels in which are all belongs to China Ⅲ. China Ⅲ is the only group enrolled in the world range epidemical circulating, having a close relation with the strains from Mongolia state and Russia.Reports of rabies cases in Qinghai province and Xizang province are rarely seen for decades,until the first relative cases was found in 2012 and 2015. Data indicates the very few found epidemic strains in Inner Mongolia belongs to China IV group that had ever found before. The China V group is up to now including only the strains ever found in the early 1990s, never played a role in the third epidemic peak that broke after 1990. China VI popularized within only Yunnan and Guangxi province. Surveillance data indicates the first Input and confirmed rabies virus, though it was proved co-belongs to a main class with China Ⅳ that has ever circulated within a limited range. Besides, the obvious differences are also worth noted. Comparing with those relative strains popularized internationally, this case had a close relation with Indian I141 and RV61 strain, belongs to Arctic-like-1 group, a widely spread group of Arctic related strains in Middle East and South Asia.Patient fund in Beijing had ever been bitten by a monkey in Indian before. Phylogenetic analysis latter demonstrated The source of infection is indeed an India wild animal. Further test had indicated this case as input-rabies. Thus, a whole genome sequence had acquired and genome sequence characteristics of different populations in China were compared and analyzed.Charpter3. Establishment of rapid detection technology for the LYSSA related virusImprove our ability in comprehensive preventing and control of emerging infectious diseases, achieve rapid pathogen screening within 2 hours. Establishment of TaqMan PCR detection method for the flanders virus （FLAV）. Primers and probes work well. The coefficients of variation of Ct values were all less than 1.7% in same sample, sensitivity of quantitative analysis model reached 100 copies/PCR. Establishment of TaqMan PCR detection method for the Tahyna virus （TAHV）, has good sensitivity, specificity and stability. The coefficients of variation of Ct values were all less than 5% in same sample, sensitivity of quantitative analysis model reached 10 copies/PCR... |
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