RT-nPCR Assays for Amplification and Sequencing of VP1 Genes in Human Enterovirus A-D from Clinical Specimens.

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 per μL and copies per μL, and the newly established methods were tested in clinical specimens collected in recent years.

Incidence of human rabies and characterization of rabies virus nucleoprotein gene in dogs in Fujian Province, Southeast China, 2002-2012.

Background: Rabies is a global fatal infectious viral disease that is characterized by a high mortality after onset of clinical symptoms. Recently, there has been an increase in the incidence of rabies in China. The aim of this study was to investigate the incidence of human rabies and characterize the rabies virus nucleoprotein gene in dogs sampled from Fujian Province, Southeast China from 2002 to 2012.

[Molecular epidemiology of HFMD-associated pathogen coxsackievirus A6 in Fujian Province, 2011-2013].

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.

Neutralizing antibodies respond to a bivalent dengue DNA vaccine or/and a recombinant bivalent antigen.

There is currently no effective vaccine to prevent dengue infection, despite the existence of multiple studies on potential methods of immunization. The aim of the present study was to explore the effect of DNA and/or recombinant protein on levels of neutralizing antibodies. For this purpose, envelope domain IIIs of dengue serotypes 1 and 2 (DEN-1/2)were spliced by a linker (Gly‑Gly‑Ser‑Gly‑Ser)3 and cloned into the prokaryotic expression plasmid pET30a (+) and eukaryotic vector pcDNA3.

The effect of temperature on the extrinsic incubation period and infection rate of dengue virus serotype 2 infection in Aedes albopictus.

Dengue fever is an acute mosquito-borne viral disease caused by dengue virus (DENV). Temperature may affect the efficiency of the mosquito vectors in spreading DENV. Aedes albopictus mosquitoes were infected orally with a DENV2 suspension and incubated at different temperatures.

[Characteristics of complete genome of pandemic A/H1N1/2009 influenza virus isolated in Fujian Province, China].

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.

[Identification of origins of marker chromosomes using fluorescence in situ hybridization].

Objective: To assess the value of fluorescence in situ hybridization (FISH) and bacterial artificial chromosome FISH (BAC-FISH) for the diagnosis for patients with marker chromosomes.

Methods: Sixteen patients with marker chromosomes were analyzed with technologies including GTG-banding, Q-banding, multiplex FISH and BAC-FISH.

Results: The marker chromosomes in the 16 patients were verified as der(Y) (2 cases), psu dic(Y) (1 case), psu dic(15) (1 case), dic(15) (1 case), del(Y) (1 case), r(X) (5 cases), i(14 or 22) (2 cases), i(18) (1 case).

Molecular epidemiology of Echovirus 30 in Fujian, China between 2001 and 2011.

Echovirus 30 (E-30) was responsible for an outbreak of aseptic meningitis between April 1 and June 2, 2011 in Fujian Province, China. A molecular epidemiology study of 115 E-30 strains was performed to characterize the genetic features of the etiologic agent of the 2011 aseptic meningitis outbreak. The phylogenetic trees of the complete VP1 gene (876 bp) from 74 of 115 isolates and 50 reference sequences were analyzed.

Molecular epidemiology of hepatitis C virus among different groups of people in the province of Fujian, China.

Genotyping of hepatitis C virus (HCV) can provide valuable information for prognosis and treatment duration prediction. To explore the genetic diversity of HCV in Fujian Province, China, 112, 104 and 48 anti-HCV-positive serum samples were collected from volunteer blood donors, IDUs and patients, respectively, from Jan 2008 to Dec 2008 and were genotyped through sequence analysis, followed by phylogenetic analysis in the C/E1 and NS5B regions. Genotypes could be determined for 85.

[Genotype I Japanese encephalitis virus is the main genotype in mosquito in Fujian province].

Objective: To grasp the infection rate and genotypes of Japanese encephalitis virus (JEV) in mosquito in Fujian province.

Methods: 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.

[Construction of sub-unit dengue vaccines and analysis of its immunogenicity].

Objective: To construct sub-unit vaccines of dengue virus type 1 to 4 and to analyze its immunogenicity.

Methods: 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.

[Sequencing and analysis of the complete genome sequence of WU polyomavirus in Fuzhou, China].

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).

Construction and characterization of a full-length infectious clone from a fast-replicating, X4-tropic HIV-1 subtype B' isolate.

In HIV-1 epidemics in China, HIV-1 subtype B' is the most predominant subtype circulating in intravenous drug users. In this study, we constructed an HIV-1 full-length infectious molecular clone based on the primary virus LWJ, which was isolated from an HIV-infected patient in Fujian Province, China. Phylogenetic and bootscanning analysis of the viral sequence revealed that the isolate LWJ belonged to HIV-1 subtype B'.

[A duplex nested PCR assay detecting of Vibrio cholerae and its application on environmental specimens].

Objective: 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.

Methods: 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.

Sharp object injuries among health care workers in a Chinese province.

Health care workers in nine hospitals in Fujian were surveyed between December 2005 and February 2006 regarding the occurrence of sharp object injuries (SOIs). Survey results indicated that 71.3% of the health care workers had sustained SOIs during the past year.

[Application of purified recombinant outer membrane lipoprotein LipL32 in detecting antibodies among leptospirosis cases].

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.

[Study on the public health emergent events in Fujian province in 2004--2007].

Objective: 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 according to the internet-based surveillance reports.

Results: From 2004 to 2007, there were 304 emergency events being surveyed.

[RtL164V, a mutation possibly associated with lamivudine resistant HBV].

Objective: To study the efficacy of lamivudine treatment in chronic hepatitis B patients affected by structures of HBV P-genes.

Methods: 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.

[A sero-epidemiology study on hepatitis E virus infection in Fujian province].

Objective: To understand the sero-prevalence of hepatitis E virus (HEV) infection among different populations and animals in Fujian province.

Methods: 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.

High-level expression of recombinant dengue virus type 2 envelope domain III protein and induction of neutralizing antibodies in BALB/C mice.

Dengue fever is a growing public health problem in many countries since so far no effective vaccines are available. In this study, the domain III of dengue virus type 2 envelope was expressed in Escherichia coli without fusion of any carrier protein. The recombinant protein was detected in the form of inclusion bodies, which were solubilized in 8M urea and could be purified subsequently by high-performance liquid chromatography (HPLC) on an ion exchange column.

Genetic characterization of CRF01_AE full-length human immunodeficiency virus type 1 sequences from Fujian, China.

HIV-1 CRF01_AE is the main prevalent HIV-1 recombinant strain in China and the dominant strain in Fujian as well; therefore, obtaining and understanding the genetic characterization of this clade become important. To date only a few full-length HIV-1 CRF01_AE genomes from China have been sequenced. We reported 13 full-length HIV-1 CRF01_AE isolate sequences from Fujian, China.

[Sequence analysis of the full-length glycoprotein 120 gene of HIV-1 CRF01_AE in Fujian, China].

Objective: To characterize full length glycoprotein 120 gene variations of 21 HIV-1 CRF01_AE isolated in Fujian, China, so as to help in the immunogenic research and vaccine design.

Methods: Twenty-one peripheral blood samples were randomly collected form 100 HIV-1CRF01_AE infected persons in Fujian 2004 approximately 2005. DNA was extracted, Nested-PCR was used to amplify the envelop protein full length gp120 gene.

Genetic characterization of three CRF01_AE full-length HIV type 1 sequences from Fujian Province, China.

Background: 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.