RT-nPCR Assays for Amplification and Sequencing of VP1 Genes in Human Enterovirus A-D from Clinical Specimens / 生物医学与环境科学（英文）

Objective@#To develop RT-nPCR assays for amplifying partial and complete VP1 genes of human enteroviruses (HEVs) from clinical samples and to contribute to etiological surveillance of HEV-related diseases.@*Methods@#A panel of RT-nPCR assays, consisting of published combined primer pairs for VP1 genes of HEV A-C and in-house designed primers for HEV-D, was established in this study. The sensitivity of each RT-nPCR assay was evaluated with serially diluted virus stocks of five serotypes expressed as CCID @*Results@#The sensitivity of RT-nPCR assays for amplifying partial VP1 gene of HEVs was 0.1 CCID @*Conclusion@#This RT-nPCR system is capable of amplifying the partial and complete VP1 gene of HEV A-D, providing rapid, sensitive, and reliable options for molecular typing and molecular epidemiology of HEVs in clinical specimens.

Bivariate heritability estimation of resting heart rate and common chronic disease based on extended pedigrees / 北京大学学报(医学版)

OBJECTIVE@#To estimate the univariate heritability of resting heart rate and common chronic disease such as hypertension, diabetes, and dyslipidemia based on extended pedigrees in Fujian Tulou area and to explore bivariate heritability to test for the genetic correlation between resting heart rate and other relative phenotypes.@*METHODS@#The study was conducted in Tulou area of Nanjing County, Fujian Province from August 2015 to December 2017. The participants were residents with Zhang surname and their relatives from Taxia Village, Qujiang Village, and Nanou Village or residents with Chen surname and their relatives from Caoban Village, Tumei Village, and Beiling Village. The baseline survey recruited 1 563 family members from 452 extended pedigrees. The pedigree reconstruction was based on the family information registration and the genealogy booklet. Univariate and bivariate heritability was estimated using variance component models for continuous variables, and susceptibility-threshold model for binary variables.@*RESULTS@#The pedigree reconstruction identified 1 seven-generation pedigree, 2 five-generation pedigrees, 23 four-generation pedigrees, 186 three-generation pedigrees, and 240 two-generation pedigrees. The mean age of the participants was 57.2 years and the males accounted for 39.4%. The prevalence of hypertension, diabetes, dyslipidemia in this population was 49.2%, 10.0%, and 45.2%, respectively. The univariate heritability estimation of resting heart rate, hypertension, and dyslipidemia was 0.263 (95%CI: 0.120－0.407), 0.404 (95%CI: 0.135－0.673), and 0.799 (95%CI: 0.590－1), respectively. The heritability of systolic blood pressure, diastolic blood pressure, fasting glucose, total cholesterol, triglyceride, high-density lipoprotein cholesterol, and low-density lipoprotein cholesterol was 0.379, 0.306, 0.393, 0.452, 0.568, 0.852, and 0.387, respectively. In bivariate analysis, there were phenotypic correlations between resting heart rate with hypertension, diabetes, diastolic blood pressure, fasting glucose, and triglyceride. After taking resting heart rate into account, there were strong genetic correlations between resting heart rate with fasting glucose (genetic correlation 0.485, 95%CI: 0.120－1, P<0.05) and diabetes (genetic correlation 0.795, 95%CI: 0.181－0.788, P<0.05).@*CONCLUSION@#Resting heart rate was a heritable trait and correlated with several common chronic diseases and related traits. There was strong genetic correlation between resting heart rate with fasting glucose and diabetes, suggesting that they may share common genetic risk factors.

Elimination of latent HIV genome and its infected cells by CRISPR/Cas9 system / 中国人兽共患病学报

The incidence and mortality of AIDS have been decreasing after the adoption of combined antiretroviral therapy strategy in the world,then AIDS has become a manageable chronic infectious disease.But HIV/AIDS continues to be a major global public health problem since it is restricted by a variety of factors.The major reason for the persistence of HIV/AIDS is the inability of existing treatments to clear or eradicate the multiple HIV reservoirs that exist in the human body.To suppress the virus replication and rebound,HIV/AIDS patients must take life-long antiviral medications.A few years ago,the clustered regularly interspaced palindromic repeats (CRISPR)/CRISPR-associated nuclease 9 (Cas9)system has been developed as a simple,fast and easy to operate gene-editing technique.Several studies in HIV infected cells and/or in animal models have shown that the system has the potential to eliminate or disrupt HIV-integrated genome or HIV-infected cells from multiple HIV reservoirs,which may result in the complete cure of HIV/AIDS.This paper analyzes the results of CRISPR/CAS9 in the elimination of latent HIV,and discusses the possible problems and trends.

Research progress of HIV-1 drug resistance mutations / 中国人兽共患病学报

By the end of 2015,all over the world there were around 17 million HIV/AIDS cases received antiretroviral therapy,the HIV-1 morbidity and mortality decreased rapidly.With antiretroviral treatment to all HIV infected persons,HIV resistance mutation is also a threat to the long-term treatment and also,had a negative impact on the important public health strategy of the global elimination in 2030.This review attempts to proceed from different economic and geographical environment,describing genetic barrier of commonly used antiretroviral drugs,the degree of their cross-reactions,and the epidemiology and management of drug-resistant mutations from the individual and group levels.The paper also summarizes the prevalent modes of transmitted drug resistance (TDR) and acquired drug resistance (ADR) in both high-income and low-and middle-income countries (LMICs),and analyze the two kind problems of public health significance to HIV resistant mutations,i.e.pretreatment resistance (PDR) and preexposure prophylaxis (PREP).In addition,in view of effectivel HIV cases of treatment and management in different countries,this paper also analyzes the genotypic resistance testing and treatment practices related to AIDS prevention and control.The content has a certain reference function to our country.

Bibliometric analysis of global research production in Zika virus disease based on Web of Science / 中国人兽共患病学报

This paper described global output of scientific publications on Zika virus disease during 1990 to 2016.Based on its literature search and analysis tools integrated on the Web of Science,we analyzed the year-,country-,affiliation-and author-specific literature production,publication journals,and scopes and citation of publications.A total of 1 818 publications associated with Zika virus disease were retrieved in Web of Science,during the period from 1990 through 2016,92.68％ of the literatures were published in 2016.The 41.09 ％ of the literatures were published as "Original Article".The top 3 productive countries were USA,Brazil and France,and the top 3 productive affiliations were US CDC,Universidade de Sāo Paulo at Brazil and the University of Texas Medical Branch at Galveston in USA.The top 3 productive authors were Wiwanitkit Viroj,Musso Didier and Jamieson Denise J,and the most 3 cited authors were Musso Didier,Fischer Marc and Jamieson Denise J.A total of 111 publications by Chinese researchers were retrieved,ranking the fifth around the world,with the total citation of 727,with 6.5 citations per paper.From 1990 to 2016,American,Brazilian and French scientists contribute a lot to the research on Zika virus disease,which plays a very important role in promoting scientific development and academic communication.Chinese scientists also made contributions on Zika virus research in the science community during the past two years.

Advances in the study of hepatitis E virus transmission / 中国人兽共患病学报

Hepatitis E virus is an RNA virus,which is classified into 4 genotypes.HEV-1 and HEV-2 genotypes are transmitted by the contaminated water in human infections.HEV-3 and HEV-4 genotypes are typical zoonosis viruses,which mainly infect pig,wild boar and deer,and also could infect rabbit,rat,sheep,poultry and other animals.In addition,HEV-1 and HEV-2 oceasionally infect human beings via foodborne transmission by the contaminated water,undercooked meat and viscera.HEV Infection is a self limiting disease,but a few of the infections turn into chronic hepatitis,cirrhosis and other liver diseases.HEV infection is globally prevelent.In China,the majority of infections HEV-4 caused sporadic cases except an HEV-1 outbreak in Xinjiang 30 years ago.The continuous development of the "One Belt and Road" initiative may increase the risk of introducing more virulent HEV-1 viruses from overseas countries.China has developed a vaccine(HEV239) to prevent the infection of hepatitis E virus,Which will play an important role in the control of HEV epidemic in China.

Molecular epidemiology of HFMD-associated pathogen coxsackievirus A6 in Fujian Province, 2011-2013 / 病毒学报

In order to characterize the molecular epidemiology of HFMD-associated Coxsackievirus A6 (CVA6) in Fujian Province, a total of 1340 specimens from non-EV71 non-CVA16 HFMD patients were collected during 2011-2013. Isolated virus strains were identified and subtyped. Full-length coding regions for the VP1 gene of the predominant serotype CVA6 isolates were amplified and sequenced. Among the 375 non-EV71 non-CVA16 HFMD cases confirmed by virus isolation and molecular subtyping, 182 (48.5%) were found to be caused by CVA6, accounting for 7.9%, 16.2% and 39.6% HFMD-associated enteroviruses in FujianProvince during 2011, 2012, and 2013, respectively. Compared with general features observed in the HFMD epidemic, no difference in CVA6-specificity or severity rates was observed between geographical origins, gender, or age groups. Nucleotide sequence analyses of VP1 genes revealed high diversity levels of 16.2%-18.6% among CVA6 strains from Fujian Province, in contrast to the prototype CVA6 strain, and showed low levels of diversity in the amino acid sequences (4.3%-6.2%). Phylogenetic analysis also indicated that CVA6 isolates from Fujian Province were distinct from the prototype strain and other isolates from abroad; however, it was homologous to domestic strains, although the Fujian isolates clustered into multiple branches. These results suggested that significant changes in the pathogenic spectrum of HFMD in Fujian Province occurred during 2011-2013, as CVA6 was one of the predominant serotypes of HFMD. CVA6 isolates from Fujian Province were co-circulating and co-evolving with other domestic strains as multiple closely related CVA6 transmission chains were observed in Fujian Province overall and within each prefecture.

Characteristics of complete genome of pandemic A/H1N1/2009 influenza virus isolated in Fujian Province, China / 病毒学报

This study aims to investigate the characteristics of genomic variation of pandemic A/H1N1/2009 influenza virus isolated in Fujian Province, China. Complete genome sequence analysis was performed on 14 strains of pandemic A/H1N1/2009 influenza virus isolated from Fujian during 2009-2012. All virus strains were typical low-pathogenic influenza viruses, with resistance to amantadine and sensitivity to neuraminidase inhibitors. Eight genome fragments of all strains were closely related to those of A/California/07/2009 (H1N1) vaccine strain, with > or = 98.2% homology. Compared with the vaccine strain, the influenza strains from Fujian had relatively large variation, and variation was identified at 11 amino acid sites of the HA gene of A/Fujiangulou/SWL1155/2012 strain, including 4 sites (H138R, L161I, S185T, and S203T) involved inthree antigen determinants (Ca, Sa, and Sb). In conclusion, the influenza vaccine has a satisfactory protective effect on Fujian population, but the influenza strains from Fujian in 2012 has antigenic drift compared with the vaccine strain, more attention should therefore be paid to the surveillance of mutations of pandemic A/H1N1/2009 influenza virus.

Genotype I Japanese encephalitis virus is the main genotype in mosquito in Fujian province / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To grasp the infection rate and genotypes of Japanese encephalitis virus (JEV) in mosquito in Fujian province.</p><p><b>METHODS</b>Mosquito specimens in Sanming city, Jianyang city and Fuzhou city in Fujian province were collected in 2010. RT-PCR was used to detect the JEV sequence from the mosquitoes by specific primers. The sequence splicing and the differentiation analysis for nucleotides, deduced amino acid sequence and phylogenetic tree were performed by the software of ATGC, Clustal X (1.83), MegAlign, GeneDoc 3.2 and Mega (4.0).</p><p><b>RESULTS</b>Totally 6987 mosquitoes were collected and main species was Culex tritaeniorhynchus and Anopheles sinensis. The infection rate of JEV in mosquitoes in Sanming, Jianyang and Fuzhou were 1.25%, 1.76% and 0.65%, respectively. One full genome in the positive specimens was sequenced. And further study showed that the positive JEV sequences belonged to genotype I.</p><p><b>CONCLUSION</b>Genotype I Japanese encephalitis virus is the main genotype in mosquitos in Fujian province.</p>

Sequencing and analysis of the complete genome sequence of WU polyomavirus in Fuzhou, China / 病毒学报

WU polyomavirus (WUPyV), a new member of the genus Polyomavirus in the family Polyomaviridae, is recently found in patients with respiratory tract infections. In our study, the complete genome of the two WUPyV isolates (FZ18, FZTF) were sequenced and deposited in GenBank (accession nos. FJ890981, FJ890982). The two sequences of the WUPyV isolates in this study varied little from each other. Compared with other complete genome sequences of WUPyV in GenBank (strain B0, S1-S4, CLFF, accession nos. EF444549, EF444550, EF444551, EF444552, EF444553, EU296475 respectively), the sequence length in nucleotides is 5228bp, 1bp shorter than the known sequences. The deleted base pair was at nucleotide position 4536 in the non-coding region of large T antigen (LTAg). The genome of the WUPyV encoded for five proteins. They were three capsid proteins: VP2, VP1, VP3 and LTAg, small T antigen (STAg), respectively. To investigate whether these nucleotide sequences had any unique features, we compared the genome sequence of the 2 WUPyV isolates in Fuzhou, China to those documented in the GenBank database by using PHYLIP software version 3.65 and the neighbor-joining method. The 2 WUPyV strains in our study were clustered together. Strain FZTF was more closed to the reference strain B0 of Australian than strain FZ18.

Construction of sub-unit dengue vaccines and analysis of its immunogenicity / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To construct sub-unit vaccines of dengue virus type 1 to 4 and to analyze its immunogenicity.</p><p><b>METHODS</b>Envelope domain III s of dengue serotypes 1 and 2, as well as 3 and 4, were spliced by a linker (Gly-Gly-Ser-Gly-Ser)3 and cloned into vector pET-30a, then transformed into E. coli to express recombinant fusion proteins. The recombinant proteins were purified by high-performance liquid chromatography and mixed to immunize BALB/c mice. The neutralizing antibodies were tested by neutralizing assay, as well as in newborn mice challenged intracranially with dengue virus type 1 to 4.</p><p><b>RESULTS</b>Mice immunized with proteins could produce neutralizing antibodies, with titers of 1:34. 9, 1: 45.3, 1: 24.7 and 1:38.4 for DEN-1 to 4 respectively. 100% newborn mice challenged with DEN-1 or 2 in combination with sera from mice immunized with recombinant proteins were protected, whereas 83% protection was obtained when challenged with DEN-3 or 4.</p><p><b>CONCLUSION</b>The recombinant proteins possess excellent immunogenicity to induce neutralizing antibodies and would be valuable for development of a tetravalent sub-unit vaccine.</p>

A duplex nested PCR assay detecting of Vibrio cholerae and its application on environmental specimens / 中华预防医学杂志

<p><b>OBJECTIVE</b>To establish a duplex nested PCR assay system which is capable for detecting O1 and O139 groups of Vibrio cholerae simultaneously, and is applicable to environmental specimens from routine cholera surveillance.</p><p><b>METHODS</b>Based on nucleic acid sequences available in GenBank, six sets of primers were designed by PrimerSelect program of DNAStar, targeting the rfb gene that encodes the O antigens of O1 and O139 V. cholerae, respectively. The specificity of several primer combinations was tested. A duplex nested PCR assay system for simultaneously detecting O1 and O139 V. cholerae was established, subsequently, its sensitivity, specificity, reproducibility and field evaluation were tested. The sensitivity of this assay was evaluated by comparing detection limits of nested PCR and conventional PCR. Its reproducibility was tested by 32 positive samples (11 samples positive for O1, 21 samples positive for O139) from environmental surveillance. In addition, the selected amplicons from positive samples were sequenced and analyzed with relevant sequences.</p><p><b>RESULTS</b>This newly-established duplex nested PCR assay might distinguish O1 V. cholerae from O139 V. cholerae, based on fragment lengths of amplicons, with reliable reproducibility, and no specific amplification was observed as compared with other vibrio species. The sensitivity of this nested PCR was (15 000) higher than conventional PCR, and there was no interference observed with multiple primers and complicated templates in the same vial. In its field evaluation, 32 positive DNA samples were detected and be further confirmed with double or triple tests, implying reliable reproducibility and consistency of this system. These results indicated that this assay had reliable reproducibility. No amplification was observed in all negative specimens and also suggested the acceptable specificity of this assay. Sequence analysis of the selected amplification products revealed 100% homogeneous with relevant genes from V.cholerae, indicating that these amplicons were originated from V. cholerae.</p><p><b>CONCLUSION</b>This duplex nested PCR assay system should be rapid, sensitive and especially applicable to small laboratories, and be suitable for dynamic environmental surveillance.</p>

Application of purified recombinant outer membrane lipoprotein LipL32 in detecting antibodies among leptospirosis cases / 中华流行病学杂志

Objective To establish recombinant outer membrane lipoprotein LipL32-based antibody detection assays in identifying leptospirosis. Methods Recombinant leptospiral outer membrane protein LipL32 was obtained by genetic engineering method. This purified protein was used in the indirect and sandwich ELISA assays to test the antibodies in sera of human beings and rats, and the results were compared with those obtained by microscopy agglutination test (MAT) and imported ELISA kit. Results When the acute and convalescent phase specimens from 9 leptospiral patients were tested, the detected rates of three ELISAs were similar to the MAT. Among the 45 probable cases which MAT showed positive, 32 (71.11%) samples were positive by r32-I-ELISA, 36(80.00%) by r32-S-ELISA,while 28.89% (13/45) samples were positive and 55.56% (25/45)were suspicious by D.A.I-ELISA. The specificity of r32-I-ELISA and r32-S-ELISA were 97.10 % (67/69) for 69 specimens. 43 healthy specimens were negative by both r32-I-ELISA and r32-S-ELISA, 14 healthy specimens were negative by D.A.I-ELISA. Among 16 non-leptospirosis patients, two specimens were positive by r32-I-ELISA and r32-S-ELISA, D.A.I-ELISA and identified one positive specimen, while 12 specimens were suspicious by D.A.I-ELISA. For 10 syphilis specimens, data obtained through three ELISAs were in consistent with that by MAT. A sandwiched ELISA, using rLipL32 protein as the antigen was developed to detect rat sera. Considering MAT as standard test, the sensitivity and specificity were 86.75 % (131/151), 99.19 % (122/123) respectively with coincidence rate as 92.34% (253/274). Conclusion The recombinant protein LipL32 had high immunoresctivity and could be used as an antigen for the detection of panthogenic leptospirosis. In summary, the novel sandwiched ELISA with rLipL32 showed similar sensitivity and specificity to that of MAT in the antibody detection of rat leptospirosis. It was suitable for large scales field sero-epidemiological studies.

RtL164V, a mutation possibly associated with lamivudine resistant HBV / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To study the efficacy of lamivudine treatment in chronic hepatitis B patients affected by structures of HBV P-genes.</p><p><b>METHODS</b>P genes of HBV isolated from sera were amplified by means of one-step PCR and then sequenced. The sequences of the P-genes from responders, primary non-responders and rebounders were compared before and after their lamivudine treatments.</p><p><b>RESULTS</b>(1) Among the primary non-responders and rebounders, commixture genotype B and C was found in 2 patients with genotype B and in 1 patient with genotype C; genotype shift from B into C was also observed in one patient after lamivudine therapy. (2) During the course of the therapy YMDD mutation emerged in all 8 primary non-responders and rebounders, which existed in some patients of the 3 groups before their lamivudine treatment. (3) An rtL164V mutation in the reverse transcriptase region was observed in all primary non-responders before and after lamivudine therapy and also in rebounders when viral breakthrough occurred, which was not seen in the responders. (4) Four amino acid substitutions at rt91, rt168, rt234 and rt256 in the reverse transcriptase region were seen in the rebounders and primary non-responders.</p><p><b>CONCLUSION</b>YMDD mutation was not the only key point closely linked to HBV resistant to lamivudine therapy. RtL164V may be a novel mutation correlated with lamivudine-resistance.</p>

Study on the public health emergent events in Fujian province in 2004-2007 / 中华流行病学杂志

Objeetive To study the public health emergent events(PHEE)in Fujian province，from 2004 to 2007.Methods Descriptive and analytic methods were Used to analyze the PHEE in Fujian province aecording to the internet.based surveillance reports.Results From 2004 to 2007.there were 304 emergency events being surveyed.Of all the events，there were 7(2.30％)belonged to serious-degree of grade II，57(18.75％)to gradeⅢand 240(78.95％)t0 gradeⅣ，but with no grade I.Results showed that the attack rate in affected population WaS 25.82‰.the mortality rate was 0.08‰and the fatalky rate Was 0.32％.The numbers of emergency events decreased 2.82％on average.each year.A total number of 169(55.60％)events occurred in schools with 71(23.36％)in the countryside.Numbers due to infectious disease-born Was 233(76.64％)including avian flu，cholera and dengue fever were predominant pathogens of the grade II and grade emergency events.57(18.75％)of the events was due to food poisoning.The epi.garph showed that there were two peaks.I.e.in Mar-Apr and Sep.contributed 43.1％to the total number of events.Conclusion Emergency events showed a stable decrease in FujJan province with communicable disease and food poisoning the two major sources and more commonly seen in schools and countryside.We suggest that the government and community pay more attention to the emergency events of avian flu,cholera and dengue fever.

A sero-epidemiology study on hepatitis E virus infection in Fujian province / 中华流行病学杂志

<p><b>OBJECTIVE</b>To understand the sero-prevalence of hepatitis E virus (HEV) infection among different populations and animals in Fujian province.</p><p><b>METHODS</b>One thousand one hundred and fifty-one serum samples were collected from 5 species of animals including swine, dog, cow, sheep and rat. A total of 2209 and 1722 serum samples from the general population and from the exposed population were collected. Anti-HEV IgG was detected by ELISA. The general population was composed of healthy blood donors and the individuals who had attended physical examination including farmers, handlers, veterinarians, cooks who worked with pigs or chickens while the poultry wholesale suppliers made up the exposure population.</p><p><b>RESULTS</b>The infection rates of HEV in animals were different between species (chi2 = 406.25, P < 0.01) with the highest seen in the pig group. With pigs being kept at home, the rates were between 70.00% and 94.12% but the rate was 39.77% for those families that keeping the pigs at farms. The infection rate of HEV was 23.3% in the general population and 33.3% in the exposed populations, respectively. A significantly higher infection rate for anti-HEV was found in the exposed population when comparing with general population. The positive rate of anti-HEV IgG was significantly higher in the exposed population that closely having had contact with chickens than those who had contact with pigs. The increasing trend of HEV infection rate with age had been found but there was no significant difference between males and females in the general population. In the exposed population, the infection rate in males was significantly higher than that in females.</p><p><b>CONCLUSION</b>The infection ratse of HEV in pigs and in the exposure population were much higher, especially for those persons in close contact with chickens or pigs, suggesting that the sub-clinical infection for HEV might exist. These data further supported the hypothesis that HEV might have been an zoonotic disease.</p>

Study on the epidemiology and etiologic agent of Dengue fever outbreaks in Fuzhou in 2004 / 中华流行病学杂志

<p><b>OBJECTIVE</b>To study the epidemiology and etiologic characteristics of a Dengue fever outbreak in Fuzhou from the beginning of September to the end of October in 2004 in order to understand the source of infection.</p><p><b>METHODS</b>Data on descriptive epidemiology was collected to study the characteristics and related factors to the epidemic. Dengue virus was isolated through the use of C6/36 cell line while viral serotypes were identified by indirect immunofluorecent assay with type-specific monoclonal antibody. The sources of infection were traced by nucleotide sequencing.</p><p><b>RESULTS</b>During the epidemic, 93 cases occured consistently with the region entomoplily growth and decay. The viruses of 6 strains isolated from 10 patients' blood specimens were identified as dengue virus type 1. Phylogenetic evidence suggested that the viral isolate had high genetic relation with the isolates from Kampuchea (DENV-1/KHM/2001; GenBank Accession No. L0904278).</p><p><b>CONCLUSION</b>The epidemic was caused by introduction of patients migrating into Fuzhou.</p>

Molecular identification and genetic analysis of an echovirus 19-associated epidemic of aseptic encephalitis / 中华流行病学杂志

<p><b>OBJECTIVE</b>To ascertain the pathogen of aseptic encephalitis epidemic in Long-Yan city in Fujian, and to find out the genetic characteristics of the virus.</p><p><b>METHODS</b>Rapid detection of enteroviral RNA by reverse transcription polymerasechain reaction (RT-PCR) was directly carried out in cerebrospinal fluid(CSF) to isolate and identify the viruses from CSF at the same time, and to detect the neutralization antibody in two serum specimens collected in acute and convalescence phase. Nucleotides of VP1 region was also analyzed by constructing phylogenetic tree.</p><p><b>RESULTS</b>ECHO 19 infection was rapidly diagnosed and sequence analysed by RT-PCR, and then echovirus type 19 from 16 of 30 CSF samples (53.33%) was isolated and detected using RD and Hep-2 cells simultaneity. The titer of ECHO 19 neutralization antibody became positive or increased by 4 times from acute to convalescence phase in 4 of the 5 patients. Phylogenetic analyses of the VP1 genes of these isolates showed that their nucleotides identity were 98.9% -100.0% which were different from those ECHO 19 from GeneBank database by 13.0%-22.4%.</p><p><b>CONCLUSION</b>The etiology of the epidemic of aseptic encephalitis was attributed to ECHO 19. The method of molecular identification not only provided rapid diagnosis of enterovirus infections, but also information about the genetic character of the viruses.</p>

Genetic characterization of three CRF01_AE full-length HIV type 1 sequences from Fujian Province, China / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>One of the major characteristics of the human immunodeficiency virus type 1 (HIV-1) is its unusually high degree of genetic variability, which involves in genetic diagnosis, subtyping, vaccine design, and epidemiology. HIV-1 CRF01_AE is a main prevalent HIV-1 recombinant strain in China. In this study, three full-length CRF01_AE genomes from Fujian Province, China were cloned, sequenced, and analyzed; and the further genetic diversity defining and epidemiologic analysis were carried out.</p><p><b>METHODS</b>Proviral DNA was extracted from non-cultured peripheral blood mononuclear cells, the near full-length HIV-1 genome was amplified and the PCR products were cloned into pCR-XL-TOPO vector and sequenced. 5'-long terminal repeat (LTR) and 3'-LTRs were amplified by additional independent PCR and cloned into pMD18T vector. Gene-based phylogenic tree was constructed and genetic distances were calculated by MEGA 3.1. Simplot was used for Bootscan analysis.</p><p><b>RESULTS</b>The phylogeny and genetic distance analysis of the three near full-length sequences confirmed that these three samples clustered with CRF01_AE isolates, more close to Thailand CRF01_AE strain CM240, and were distantly related to African CRF01_AE strain 90CF402. Analysis of their genomic organization revealed the presence of nine potential open reading frames. There were no major deletions, rearrangements, or insertions in the three sequences, but an in-frame stop codon was found in tat gene of Fj051. LTRs of the three sequences contained a few nucleotides mutation. We did not find new mosaic recombinant in the three sequences. The V3 motif was GPGQ in all the three sequences, and there were only few amino acids differences in all three V3 loop sequences.</p><p><b>CONCLUSION</b>This report reveals the background of the three full-length CRF01_AE genomes, the most dominantly circulating HIV-1 strain in Fujian Province, China. The work is essential for the design and development of an effective AIDS vaccine for the region.</p>

Sero-epidemiological study on the human T-cell leukaemia virus type I/II infection in the east coastal areas of Fujian province / 中华流行病学杂志

<p><b>OBJECTIVE</b>To study the seroprevalence of human T-cell leukaemia virus type I/II (HTLV-I/II) infection in adult population in the east coastal areas of Fujian and to explore the possible risk factors of HTLV-I/II.</p><p><b>METHODS</b>A total number of 3259 blood samples from drug users, sexually transmitted disease (STD) patients, prostitutes and blood donors for serologic assays during 1999 to 2002, were collected. All samples were screened for HTLV-I/II antibody, using enzyme linked immunosorbent assay (ELISA) kits. All of the positive samples were confirmed by western blot (WB) kits. Statistical analysis was done by Epi software, and chi(2) test by Fisher's exact test. P value < 0.05 was considered statistically significant.</p><p><b>RESULTS</b>The overall seroprevalence rate of HTLV-I/II in healthy populations was 0.06% including, 0.32% in drug users, 0.58% in STD patients and prostitutes respectively. HTLV-II had not been found. The seropositive rates for HTLV-I in STD patients and prostitutes were significantly higher than the findings among healthy populations (P < 0.05). There were no different seroprevalence rates between drug users and healthy populations (P > 0.05). No significant changes in HTLV-I prevalence rates were found in the different age groups as well as in Fuzhou and Linde cities (P > 0.05).</p><p><b>CONCLUSION</b>The result suggested that in the east coastal areas of Fujian province, HTLV-I was the main prevalent virus. The seroprevalence of HTLV-I was very low, with no HTLV-II. Neither age nor gender seemed to be HTLV-I risk factor in the east coastal areas of Fujian province, but the increase of exposure to sex might be one.</p>