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1.
Article in Chinese | WPRIM | ID: wpr-885658

ABSTRACT

Objective:To evaluate the immunological efficacy of a novel DNA vaccine against West Nile virus (WNV) in a mouse model.Methods:A DNA vaccine VRC-prME expressing the precursor membrane (prM) and envelope protein (E) of WNV Xinjiang strain (XJ11129-3) was constructed and its ability to express virus-like particles was verified in vitro. C57BL/6 mice were immunized twice with VRC-prME via intramuscular injection combined with electroporation with an interval of four weeks. Enzyme-linked immunoassay (ELISA) was used to detect serum antibodies after immunization. WNV (NY99 strain) single-round infectious particles were used to detect neutralizing antibodies. Cellular immune responses were analyzed by enzyme-linked immunoblot assay (ELISPOT) and intracellular cytokine staining (ICS). Results:VRC-prME induced a strong Th1-biased antibody response in mice that could cross-neutralize the WNV (NY99 strain) single-round infectious particles two weeks after the boost immunization. Moreover, the vaccine also elicited antigen-specific multifunctional CD8 + T cell responses (IFN-γ, IL-2, TNF-α). Conclusions:The novel DNA vaccine prepared in this study, expressing the prME protein of WNV XJ11129-3 strain, could induce stronger humoral and cellular immune responses in mice, which was worthy of further research and development for the prevention of WNV infection in China.

2.
Article in Chinese | WPRIM | ID: wpr-805153

ABSTRACT

Objective@#To understand the situation of ticks carrying pathogens in border areas of Heilongjiang province.@*Methods@#From 2009 to 2018, tick specimens were collected in Yichun, Daxing′anling area and Jiamusi in Heilongjiang province. A total of 2 530 ticks were studied, including 800 Ixodes persulcatus and 1 730 Dermacentor silvarum. Tick-borne encephalitis virus (TBEV), severe fever with thrombocytopenia syndrome virus (SFTSV), omsk hemorrhagic fever virus (OHFV), langat virus (LGTV), powassan virus (POWV) were detected by real-time RT-PCR. Spotted fever group rickettsia (SFGR) and Borrelia burgdorferi sensulato (B.b.s.l) were detected by PCR in ticks collected from Jiamusi area.@*Results@#All tick speciments collected were negative for TBEV, SFTSV, OHFV, LGTV and POWV. Tick specimens from Jiamusi carried SFGR and B. b.s.l.with positive rates of 59.5% and 8.9%.@*Conclusions@#The ticks in border areas of Heilongjiang province carry spotted fever group rickettsia and Borrelia burgdorferi, and the carrying rate of spotted fever group rickettsia is high. The monitoring and control of ticks and tick-borne diseases should be strengthened.

3.
Article in Chinese | WPRIM | ID: wpr-804826

ABSTRACT

Objective@#To investigate the west nile virus (WNV) infection in Xinjiang, China.@*Methods@#Serum samples were collected from patients with fever and chicken in southern Xinjiang, 2012. The presence of WNV-specific immunoglobulin M (IgM) antibodies and neutralizing antibodies was examined through enzyme-linked immunosorbent assay (ELISA) and plaque reduction neutraization test (PRNT90).@*Results@#A total of 1 712 serum samples of outpatients and inpatients were collected in 8 counties in southern Xinjiang. As a result , 22 samples were positive for WNV IgM antibody and 48 samples were positive for WNV neutralization antibody, among which 21 WNV IgM antibody positive samples and 42 WNV neutralization antibody positive samples were from Jiashi county. Of 383 chicken serum samples collected in 4 counties in southern Xinjiang, only 28 samples were positive for WNV neutralizing antibody, interestingly, all positive chicken serum samples were collected from Jiashi county.@*Conclusions@#This study revealed that WNV infection occurred in human and poultry in southern Xinjiang, 2012, mainly in Jiashi county.

4.
Article in Chinese | WPRIM | ID: wpr-806647

ABSTRACT

Objective@#To analyze the epidemiological characteristics and distribution characteristics of tick-borne encephalitis in China in 2014, and to provide scientific basis for formulating specific prevention and control measures.@*Methods@#The epidemic data were obtained from the "infectious disease report information management system" , using Excel 2016, GIS and other software to summarize and analyze the cases of tick borne encephalitis (TBE) reported, using the number of cases, incidence, composition ratio and other indicators to analyze and describe the TBE epidemiological characteristics in China in 2014.@*Results@#In 2014, a total of 322 cases of TBE were reported in 9 provinces in China, with an annual incidence of 0.024/100, 000 and 1 death of patient. The provinces with high number of cases were Jilin province, Inner mongolia autonomous region and Heilongjiang province, and the number of cases in the other six provinces is no more than two. TBE was distributed in spring and summer, and it is concentrated in May to July. The age of the affected population was mostly concentrated in 40-49 years old, the male-female ratio was 1.6∶1 (198/124), and the patients were dominantly farmers, household and unemployed workers, and forestry workers, they accounted for 49.40% (159/322), 26.40% (85/322) and 18.60% (60/322) of the national TBE cases respectively. The three hospitals that reported the most TBE cases in 2014 were Inner mongolia forestry general hospital, Jiangyuan People′s hospital of Baishan city, Jilin province and Mudanjiang forestry central hospital of Heilongjiang province. The number of reported cases in these three hospitals accounted for 68.6% of the whole country. The laboratory diagnosis rate of Inner mongolia forestry general hospital was the highest (91.9%).@*Conclusions@#In 2014, the incidence of TBE in China has continued to rise compared with the previous two years. The geographical focus is mainly on the forest areas of Daxing′anling, Xiaoxing′anling and Changbai Mountain.

5.
Article in Chinese | WPRIM | ID: wpr-805909

ABSTRACT

Objective@#To analysis the genotype of Japanese encephalitis virus (JEV) in mosquitoes from Shandong province.@*Methods@#Mosquitoes were collected between August and September in Weishan county, Junan county, and Kenli county of Shandong province in 2016. Viruses were isolated by BHK-21 cell and identified by molecular method . Real-Time RT-PCR was conducted to detect the Japanese encephalitis virus carried by the mosquitoes.@*Results@#A total of 8418 mosquitoes divided into 81 pools including 3 species, Culex tritaeniorhynchus, Anopheles sinensis and Armigeres obturbans. Eight Japanese encephalitis viruses were isolated; 23 pools were positive by JEV specific real-time RT-PCR. Phylogenetic analysis on E sequence of JEV showed all JEV strains belonged to genotype Ⅰ JEV, and new strains that were homogenous with previous JEV strains isolated from Shandong.@*Conclusions@#Genotype Ⅰ JEV was the dominant genotype in Shandong province.

6.
Chinese Journal of Epidemiology ; (12): 1381-1386, 2018.
Article in Chinese | WPRIM | ID: wpr-738156

ABSTRACT

Objective To investigate the distribution patterns of mosquitoes,midges and related arboviruses in Sichuan province.Methods Blood-sucking insects were collected from houses and pens,using the ultraviolet lights.Mosquito samples were classified according to morphologic characteristics and then stored at liquid nitrogen.All samples were incubated with BHK-21 and C6/36 cells for virus isolation and then detected for their viral genes.Sequences of the virus were identified and analyzed by molecular biological software,such as BioEdit 7.0.5.3,MEGA 6.0.Results In total,17 019 mosquitoes from 3 genera and 4 species and 12 700 midges were collected from the southeast regions of Sichuan province in 2016 and 2017.Among them,79.4% (13 519/17 019) belonged to Culex tritaeniorhynchus with 11.1% (1 897/17 019) as Armigeres subalbatus,5.5% (930/17 019) were Anopheles sinensis and 4.0% (673/17 019) were Anopheles sinensis 3 virus strains that isolated from Culex tritaeniorhynchus were identified as type Ⅰ Japanese encephalitis virus.Seven pools of mosquitoes isolated from Hejiang county were identified Japanese encephalitis virus gene positive through PCR amplification.With 4 pool midges were detected positive for Akabane virus through PCR gene amplification while midges samples didn't have virus isolates.Conclusions Culex tritaeniorhynchus appeared the predominant species in the southeast regions of Sichuan.Japanese encephalitis virus transmitted by mosquitoes and Akabane virus by midges were prevalent in southeast Sichuan province.

7.
Chinese Journal of Epidemiology ; (12): 1381-1386, 2018.
Article in Chinese | WPRIM | ID: wpr-736688

ABSTRACT

Objective To investigate the distribution patterns of mosquitoes,midges and related arboviruses in Sichuan province.Methods Blood-sucking insects were collected from houses and pens,using the ultraviolet lights.Mosquito samples were classified according to morphologic characteristics and then stored at liquid nitrogen.All samples were incubated with BHK-21 and C6/36 cells for virus isolation and then detected for their viral genes.Sequences of the virus were identified and analyzed by molecular biological software,such as BioEdit 7.0.5.3,MEGA 6.0.Results In total,17 019 mosquitoes from 3 genera and 4 species and 12 700 midges were collected from the southeast regions of Sichuan province in 2016 and 2017.Among them,79.4% (13 519/17 019) belonged to Culex tritaeniorhynchus with 11.1% (1 897/17 019) as Armigeres subalbatus,5.5% (930/17 019) were Anopheles sinensis and 4.0% (673/17 019) were Anopheles sinensis 3 virus strains that isolated from Culex tritaeniorhynchus were identified as type Ⅰ Japanese encephalitis virus.Seven pools of mosquitoes isolated from Hejiang county were identified Japanese encephalitis virus gene positive through PCR amplification.With 4 pool midges were detected positive for Akabane virus through PCR gene amplification while midges samples didn't have virus isolates.Conclusions Culex tritaeniorhynchus appeared the predominant species in the southeast regions of Sichuan.Japanese encephalitis virus transmitted by mosquitoes and Akabane virus by midges were prevalent in southeast Sichuan province.

8.
Article in Chinese | WPRIM | ID: wpr-808476

ABSTRACT

Objective@#To investigate the distribution patterns of arboviruses in Yunnan province near the China-Laos-Myanmar border, China, and to provide evidence for prevention and control of arboviruses diseases.@*Methods@#Mosquito samples were collected in Daluo county of Xishuangbanna Dai Autonomous Prefecture and Zhengdong county of Pu’er city in Yunnan province, 2012. Viruses were isolated from the samples by tissue culture, positive isolates were identified by RT-PCR with arbovirus species-specific primers, for further sequencing and phylogenetic analysis.@*Results@#A total of 17 species of mosquitoes from 6 genera were collected. A total of 24 strains of viruses were isolated from the mosquito pools and identified as Tembusu virus (TMUV) (2 strains), Japanese encephalitis virus (JEV) (3 strains), Getah virus (GETV) (2 strains), Banna virus (BAV) (4 strains), Densovirus (DNV) (9 strains) and Nam Dinh virus (NDiV) (3 strains).@*Conclusions@#The China-Laos-Myanmar border of Yunnan province is rich in species of mosquitoes and arboviruses.

9.
Article in Chinese | WPRIM | ID: wpr-807972

ABSTRACT

Objective@#To study the genome molecular characteristics of Getah virus (SC1210) which isolated in Sichuan province in 2012.@*Methods@#Reverse transcriptase polymerase chain reaction (RT-PCR) was used to identify the isolate and the genome was sequenced by the second Ion Torrent PGM. Computer softwares, including Mega Align and Mega 6, were used to analyze the nucleotide and deduced amino acid sequence, and draw phylogenetic trees.@*Results@#SC1210 was identified as Getah virus. The full genome sequence was 11 690nt, the nucleotide and amino acid homology of the full sequence with other strains were 99.2%-99.7% and 96.5%-99.4%.The capsid protein of SC1210 consisting of 804 nucleotides, encoding 268 amino acids and the full-length of E2 protein, had 1 266 nucleotides, encoding 422 amino acids. The nucleotide homology of the capsid protein and the E2 protein with other strains were 94.9%-99.2% and 94.6%-99.6%, and the amino acid were 97%-99.6% and 97.1%-99.5%. The 3′ UTR of the virus included 402 nucleotides and there were three repeat sequence elements and 19 nucleotides conservation sequence.@*Conclusions@#The first GETV isolate SC1210 in Sichuan province has a closer relationship with Yunnan strain YN040 and a far genetic relationship with MM2021.

10.
Chinese Journal of Zoonoses ; (12): 293-299, 2017.
Article in Chinese | WPRIM | ID: wpr-610543

ABSTRACT

In order to investigate the molecular evolution and spatio-temporal migration of Getah viruses (GETV) isolated around the world,the nucleotide and deduced amino acid sequence of GETVs were analyzed and phylogenetic trees were constructed by using informatics software including ClustalX1.83,MegaAlign,GeneDOC and Mega6.0.The Bayesian Stochastic Search Variable Selection (BSSVS) program in the BEAST v 1.8.1 software package was used to analyze the spatial dynamics of the Getah virus.Results showed that the full-length of Getah virus E2 gene consists of 1 266 nueleotides,encoding 422 amino acids.And the homology of nucleotide and amino acid were 94.5% 100% and 96.4% 100% respectively.The molecular evolution analysis revealed that there were no species and geographical distribution difference existing among GETV host animals (e.g.horses and pigs) and vectors (e.g.mosquitoes).Bioinformatics analysis showed that GETV originated in Malaysia,then it was spread to Japan,China,South Korea,Mongolia,Russia,etc.GETV E2 gene was relatively stable since GETV was first isolated in 1955.The differences of species and geographical distribution did not exist among GETV host animals and vectors,and the virus has spread from tropical regions to Eurasian continent.Thus,strengthening the detection and monitoring of GETV and its infections in humans and livestock is critical.

11.
Chinese Journal of Virology ; (6): 51-57, 2015.
Article in Chinese | WPRIM | ID: wpr-280295

ABSTRACT

To evaluate the prevalence of mosquito-borne viruses in Manshi and Ruili (Yunnan Province, China), we collected 2 149 mosquitoes (17 species) in August 2010. Virus isolation was undertaken by the cul- ture of baby hamster kidney cells (BHK-21 cells). Two virus-like isolates were obtained: DHL10M117 was isolated from collected in Mangshi; DHL10M110 was obtained from Anopheles vagus collected in Rui- li. Both isolates caused cytopathic effects,illness and death in suckling mice inoculated with these isolates via the intracerebral route. Two positive amplicons, 702-bp from the S segment and 456-bp from the M segment,were obtained using reverse transcription-polymerase chain reaction using primers specific for the Akabane virus (AKV). Phylogenetic analysis suggested that these two virus stains had a distant relation- ship with AKVs from Kenya and Australia,but were genetically close to those from Japan,South Korea, and Taiwan. However,they were separate from other Asian strains and grouped into a small branch. The highest nucleotide and amino-acid sequence identity of the S segment was found with the CY-77 strain from Taiwan (96.6% and 99.6% for DHL10M117 and 96.7% and 100% for DHL10M110,respectively). Com- parison of the M segment showed they shared the highest amino acid identity with CY-77 (99.6% and 100%, respectively), whereas the highest nucleotide identity was found with the Iriki strain from Japan (99.6% and 100%, respectively). Compared with the MP496 strain from Kenya,they displayed lower lev- els of sequence homology, at 69.7% and 70.0% for nucleotide sequences of the two loci,and 91. 0% for a- mino acids. Our results identified that DHL10M117 and DHL10M110 were strains of AKV,and provided molecular biological evidence for the existence of AKV in Yunnan Province. These AKV strains that are circulating in Yunnan Province share a close genetic relationship with strains from the rest of Asia. Culex tritaeniorhynchus and Anopheles vagus may serve as transmission vectors.


Subject(s)
Amino Acid Sequence , Animals , Anopheles , Virology , Base Sequence , Bunyaviridae Infections , Virology , China , Cricetinae , Female , Humans , Insect Vectors , Virology , Male , Mice , Orthobunyavirus , Classification , Genetics , Physiology , Phylogeny , Sequence Homology , Viral Proteins , Chemistry , Genetics
12.
Chinese Journal of Virology ; (6): 264-268, 2015.
Article in Chinese | WPRIM | ID: wpr-280263

ABSTRACT

To explore the spatial distribution mechanism of Japanese encephalitis virus (JEV), PhyML v3.0 was used to build phylogenetic tree using JEV sequences in the dataset. PAUP v4.0 and Migrapyhla softz ware were then used to analyze the migration events. The results showed that a total of 95 migration events were observed during the dispersal of JEV throughout Asia. Further analysis revealed that Thailand, and several Chinese provinces (including Shandong, Shanghai, Sichuan and Yunnan), were the main migration sources of JEV. JEV spread from these migration sources as follows: from Thailand to Australia, Cambodia, Tibet and India; from Shanghai to eastern coastal Asian regions and Yunnan; from Shandong to Korea, Zhejiang, Hubei, Shanxi and Liaoning; from Sichuan mainly to inland regions of China, as well as Vietnam and Japan; and from Yunnan to Zhejiang. This study indicated that frequent migration events occurred during the dispersal of JEV in the Asia and Pacific regions, and that Thailand, Shandong, Shanghai, Sichuan and Yunnan were the sources of JEV dispersal.


Subject(s)
Asia , Epidemiology , China , Epidemiology , Encephalitis Virus, Japanese , Classification , Genetics , Physiology , Encephalitis, Japanese , Epidemiology , Virology , Phylogeny
13.
Chinese Journal of Zoonoses ; (12): 212-215, 2015.
Article in Chinese | WPRIM | ID: wpr-460505

ABSTRACT

The Flanders virus (FLAV) is a number of family Rhabdoviridae ,contains a single‐stranded ,negative‐sense vi‐ral RNA .Here we describe a molecular detection method developed for fast measurement of FLAV based on Taqman RT‐PCR method .In this study ,FLAV specific primers and probe were designed based on the FLAV L gene sequences published in GeneBank .Quantitative standard curve of FLAV TaqMan PCR was also successfully established .The specificity and stability test showed that the system is specific and the coefficient variables were all less than 1 .7% .Quantitative standard curve based on the genomic copy was drawn ,and the lowest detectable limit (LOD) of system was 100 copies/PCR ,with higher sensitivity and stability than that of the conventional RT‐PCR assay targeting the same gene .

14.
Chinese Journal of Epidemiology ; (12): 368-373, 2015.
Article in Chinese | WPRIM | ID: wpr-240092

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the species and distribution of mosquitoes and mosquito-borne arboviruses in Yuncheng city of Shanxi province, China.</p><p><b>METHODS</b>Mosquito samples were collected in 19 collection sites from Linyi county and Yongji city in Yuncheng city, in August, 2012. After identification and classification, all the specimens were homogenized and centrifuged to acquire supernatant before being inoculated to both C6/36 and BHK21 cells for viral isolation. Positive isolates were identified with arbovirus species-specific primers under RT-PCR, for further sequencing and phylogenetic analysis.</p><p><b>RESULTS</b>A total of 10 455 mosquitoes of 7 species in 4 genuese were collected. The predominant mosquito species in Linyi county was Culex pipens pallens (91.96%, 3 911/4 253), but the one in Yongji city was Culex tritaeniorhynchus (72.85%, 4 518/6 202). A total of 23 strains of viruses were isolated from the mosquito pools. 15 strains from Culex tritaeniorhynchus and Culex pipens pallens were identified as genotype I Japanese encephalitis virus (JEV). Four strains from Culex pipens pallens were identified as Culex flavivirus (CxFV). Three strains from Culex pipens pallens were identified as Culex pipiens pallens densovirus (CppDNV). One strain from Armigeres subalbatus and Aedes albopictus was identified as Getah virus (GETV).</p><p><b>CONCLUSION</b>Four kinds of arboviruses were isolated from the mosquito pools, including GETV and CxFV, which were isolated and documented in Shanxi province for the first time. In the city of Yuncheng, Culex tritaeniorhynchus had been the predominant species and major vector for transmitting JEV. Genotype I JEV remained the major JEV circulating in the local natural environment.</p>


Subject(s)
Animals , Arboviruses , Genetics , China , Cities , Culicidae , Virology , Encephalitis Virus, Japanese , Genetics , Phylogeny , Species Specificity
15.
Chinese Journal of Epidemiology ; (12): 528-532, 2014.
Article in Chinese | WPRIM | ID: wpr-737364

ABSTRACT

Objective To investigate the distribution patterns of mosquitoes and mosquito-borne viruses in Dehong prefecture,Yunnan province,China. Methods Mosquito samples were collected using the mosquito traps from five counties of Dehong prefecture on July,2007 and 2010. Mosquitoes were cell cultured for viral isolation,and positive isolates were identified using RT-PCR and sequence analysis. Results A total of 43 634 mosquitoes comprised of 29 species representing six genera were collected. Culex tritaeriorhynchus and Anopheles sinensis comprised 78.69% and 14.77% of the total. Six strains of viruses were isolated from the mosquito pools. RT-PCR and phylogenetic analysis revealed three strains from Cx. tritaeriorhynchus,identified as genotypeⅠJapanese encephalitis virus(JEV). One strain was identified from Cx. tritaeriorhynchus,as Getah virus (GETV). Two strains isolated from Cx. tritaeriorhynchus and Anopheles vagus were identified as Culex pipiens pallens Densovirus(CppDNV). Conclusion Cx. tritaeriorhynchus had been the major species of mosquitoes and mainly transmitting vector of mosquito-borne viruses in Dehong prefecture. GenotypeⅠJEV,GETV and CppDNV were the vectors causing transmission of mosquitoe-borne diseases in this area. Data from phylogenetic analysis showed that these newly discovered isolates seemed to have had close relationship with those viruses previously circulating in Yunnan and other provinces of China.

16.
Chinese Journal of Epidemiology ; (12): 528-532, 2014.
Article in Chinese | WPRIM | ID: wpr-735896

ABSTRACT

Objective To investigate the distribution patterns of mosquitoes and mosquito-borne viruses in Dehong prefecture,Yunnan province,China. Methods Mosquito samples were collected using the mosquito traps from five counties of Dehong prefecture on July,2007 and 2010. Mosquitoes were cell cultured for viral isolation,and positive isolates were identified using RT-PCR and sequence analysis. Results A total of 43 634 mosquitoes comprised of 29 species representing six genera were collected. Culex tritaeriorhynchus and Anopheles sinensis comprised 78.69% and 14.77% of the total. Six strains of viruses were isolated from the mosquito pools. RT-PCR and phylogenetic analysis revealed three strains from Cx. tritaeriorhynchus,identified as genotypeⅠJapanese encephalitis virus(JEV). One strain was identified from Cx. tritaeriorhynchus,as Getah virus (GETV). Two strains isolated from Cx. tritaeriorhynchus and Anopheles vagus were identified as Culex pipiens pallens Densovirus(CppDNV). Conclusion Cx. tritaeriorhynchus had been the major species of mosquitoes and mainly transmitting vector of mosquito-borne viruses in Dehong prefecture. GenotypeⅠJEV,GETV and CppDNV were the vectors causing transmission of mosquitoe-borne diseases in this area. Data from phylogenetic analysis showed that these newly discovered isolates seemed to have had close relationship with those viruses previously circulating in Yunnan and other provinces of China.

17.
Chinese Journal of Epidemiology ; (12): 528-532, 2014.
Article in Chinese | WPRIM | ID: wpr-348630

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the distribution patterns of mosquito and mosquito-borne viruses in Dehong prefecture, Yunnan province, China.</p><p><b>METHODS</b>Mosquito samples were collected using the mosquito traps from five counties of Dehong prefecture on July, 2007 and 2010. Mosquito were cell cultured for viral isolation, and positive isolates were identified using RT-PCR and sequence analysis.</p><p><b>RESULTS</b>A total of 43 634 mosquito comprised of 29 species representing six genera were collected. Culex tritaeniorhynchus and Anopheles sinensis comprised 78.69% and 14.77% of the total. Six strains of viruses were isolated from the mosquito pools. RT-PCR and phylogenetic analysis revealed three strains from Cx. tritaeniorhynchus, identified as genotype I Japanese encephalitis virus (JEV). One strain was identified from Cx. tritaeniorhynchus, as Getah virus (GETV). Two strains isolated from Cx. tritaeniorhynchus and Anopheles vagus were identified as Culex pipiens pallens Densovirus (CppDNV).</p><p><b>CONCLUSION</b>Cx. tritaeniorhynchus had been the major species of mosquito and mainly transmitting vector of mosquito-borne viruses in Dehong prefecture. Genotype I JEV, GETV and CppDNV were the vectors causing transmission of mosquito-borne diseases in this area. Data from phylogenetic analysis showed that these newly discovered isolates seemed to have had close relationship with those viruses previously circulating in Yunnan and other provinces of China.</p>


Subject(s)
Alphavirus , Animals , Arboviruses , Classification , China , Culicidae , Virology , Disease Vectors , Classification , Encephalitis Virus, Japanese
18.
Virologica Sinica ; (6): 179-186, 2012.
Article in Chinese | WPRIM | ID: wpr-424012

ABSTRACT

Arboviruses represent a serious problem to public health and agriculture worldwide.Fast,accurate identification of the viral agents of arbovirus-associated disease is essential for epidemiological surveillance and laboratory investigation.We developed a cost-effective,rapid,and highly sensitive one-step triplex RT-PCR enzyme hybridizationassay for simultaneous detections of Japanese Encephallitis virus (JEV,Flaviviridae)Getah virus (GETV,Togaviridae),and Tahyna virus (TAHV,Bunyaviridae) using three pairs of primers to amplify three target sequences in one RT-PCR reaction.The analytical sensitivity of this assay was 1 PFU/mL for JEV,10PFU/mL for GETV,and 10 PFU/mL for TAHV.This assay is significantly more rapid and less expensive than the traditional serological detection and single RT-PCR reaction methods.When “triplex RT-PCR enzyme hybridization” was applied to 29 cerebrospinal fluid(CSF)samples that were JEV-positive by normal RT-PCR assay,all samples were strongly positive for JEV,but negative for GETV and TAHV,demonstrating a good sensitivity,specificity,and performance at CSF specimen detection.

19.
Article in Chinese | WPRIM | ID: wpr-379811

ABSTRACT

Objective To study the genome molecular characteristics of Getah virus(DY0824)which isolated in Shandong province,2008 by molecular biology methods.Methods Reverse transcriptasepolymerase chain reaction(RT-PCR)was used to amplify the structural gene and 3'UTR fragments then the RT-PCR products were inserted into PGEM-T easy to be sequenced.Computer software was used to analyze the nucleotide and deduced amino acid sequence,and draw phylogenetic trees,including Clustal X1.83 and MegaAlign and Mega4.Results The capsid protein of DY0824 consists of 804 nucleotides,encoding 268 amino acids and the full-length of E2 protein is 1266 nucleotides,encoding 422 amino acids.The nucleotide homology of the capsid protein and the E2 protein with other strains were 95.4%-99.9%and 94.8%-99.5%,and the amino acid were 97.4%-100%and 97.6%-100%.The 3'UTR of the virus include 401 nucleotides and there are three repeat sequence elements.Conclusion Compared with the prototype virus,the Getah virus isolated in Shandong province had 7 amino acid differences in capsid protein genes and 10 amino acid differences in E protein genes.The 3'UTR region had multi-nucleotide changes.

20.
Article in Chinese | WPRIM | ID: wpr-380717

ABSTRACT

Objective To identify the Banna viruses isolated in northwestern part of Yunnan prov-ince in order to make the difference clear between the isolates and other Banna viruses isolated in other parts of Yunnan. Methods Three isolates of Banna vires isolated in 2005 and 2006 were identified by morpholo-gy, RNA-PAGE profile and molecular biologic method. Nueleotide and amino acid sequences of segment 12 of the 3 isolates were sequenced and analyzed. Results Three Banna viruses were isolated from mosquitoes collected in northwestern part of Yunnan during 2005 and 2006. Electron microscopy study showed that they are spherical with a diameter of 70 nm, no envelope but two layers of eapsid. It was found that the genome of the 3 isolates composes of 12 segments presenting band profile of 6-6 in RNA-PAGE. Nueleotide acid se-quence analysis about segment 12 showed that the identity was 99% between the 3 new isolates, 98% and 90% between the 3 isolates and the strains isolated in other parts of Yunnan, China and Indonesia, respec-tively. Phylogenetie analysis based on segment 12 gene showed that 3 new isolates clnstered in the same branch with the viruses isolated in other parts of Yunnan. The same difference of amino acids was found between Banna viruses isolated in China and Indonesia strains in the analysis of segment 12. Conclusion Banna virus strains were firstly isolated from mosquitoes collected in northwestern part of Yunnan province. Nueleotide acid sequence analysis of the 3 new isolates showed higher identity with strains isolated in other parts of Yunnan.

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