Background: While mother-to-child transmission (MTCT) of hepatitis B virus (HBV) remains a significant challenge in China, research investigating the effectiveness of the September 2017 pilot program to eliminate MTCT of HIV, syphilis, and HBV is limited. Baoan district, which has a higher-than-average rate of hepatitis B infection among pregnant women and strong support from the government, was one of six national pilot districts selected for the program. Therefore, this study aims to assess the progress and implementation of the elimination of MTCT of HBV in Baoan district over a period of 5 years.
View Article and Find Full Text PDFFleas (Order Siphonaptera) are common blood-feeding ectoparasites, which have important economic significance. Limited mitochondrial genome information has impeded the study of flea biology, population genetics and phylogenetics. The and complete mt genomes are described in this study.
View Article and Find Full Text PDFThe mitochondrial genome may include crucial data for understanding phylogenetic and molecular evolution. We sequenced the complete mitogenome of and for the first time. 's complete mitogenomes were 14,720 and 14,895 bp in size, respectively, and both contained two ribosomal RNA (rRNA) genes, 22 transfer RNA (tRNA) genes, and 13 protein-coding genes (PCG).
View Article and Find Full Text PDFTicks are deemed to be second only to mosquitoes as the most common vector of human infectious diseases worldwide that give rise to human and animal diseases and economic losses to livestock production. Our understanding of the phylogenetic analysis between tick lineages has been restricted by the phylogenetic markers of individual genes. Genomic data research could help advance our understanding of phylogenetic analysis and molecular evolution.
View Article and Find Full Text PDFTicks transmit diverse pathogens that cause human and animal diseases, leading to an increasing number of new challenges around the world. Genomic data research could help advance our learning of phylogenetic analysis and molecular evolution. Mitochondrial genome DNA has been helpful in illustrating the phylogenetic analysis of eukaryotes containing ticks.
View Article and Find Full Text PDFTo establish a MacELISA method for the detection of IgM antibodies against Chikungunya virus (CHIKV), we prepared virus like particle (VLP) antigens of CHIKV using the whole structural protein C-E3-E2-6K-E1 encoding gene with a baculovirus expression system in Sf9 insect cells. The VLPs were purified and used to immunize Kunming mice. Then, polyclonal antibodies were purified from the samples of ascites with a protein G HiTrap SP column and labeled with horseradish peroxidase.
View Article and Find Full Text PDFThis article aimed to study the antigenicity of nucleocapsid proteins (NPs) in six pathogenic phleboviruses and to provide theoretical evidence for the development of serological diagnostic reagents. NPs of six pathogenic phleboviruses were expressed and purified using a prokaryotic expression system and rabbits were immunized with individual recombinant NPs. Cross-reactions among NPs and rabbit sera were determined by both indirect ELISA and Western blotting analyses, and the sera titer was determined by indirect ELISA.
View Article and Find Full Text PDFThis study aims to investigate whether the nucleoprotein (NP) of severe fever with thrombocytopenia syndrome virus (SFTSV) can impact the cellular immunity of host cells. Gene segments that encode the NP and non-structural protein (NSs) of SFTSV were inserted into eukaryotic expression vector VR1012. Host proteins that interact with NP and affect immunity were identified with co-immunoprecipitation (IP), SDS-PAGE, mass spectrometry (MS), and Western blot.
View Article and Find Full Text PDFTo evaluate the adjuvant effect of recombinant enterovirus 71 (EV71) subunit vaccine formulated with chitosan, rabbits were orally immunized with recombinant VP1 (rVP1) or rVP1 mixed with chitosan adjuvant. Levels of virus-specific IgG and IgA antibodies in sera, mucosal wash buffer (intestine, nasal cavity, and lung), and feces were determined by indirect enzyme-linked immunosorbent assay (ELISA). The titers of neutralizing antibodies against EV71 were determined using cytopathic effect-based neutralizing assay, and levels of cytokines (IFN-gamma and IL-4) secreted from in vitro-cultured rabbit splenic lymphocytes under antigen stimulation were also determined by ELISA.
View Article and Find Full Text PDFZhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi
June 2013
Objective: To develop an assay for titration of severe fever with thrombocytopenia syndrome virus (SFTSV) based on double antibody sandwich ELISA.
Methods: A double antibody sandwich ELISA was developed for detection of SFTSV based on SFTSV nucleocapsid (N) protein specific poly- and monoclonal antibodies, procedures were optimized and evaluated. This ELISA based titration assay was compared with fluorescence assasy and plaque assay based titration method.
Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi
June 2013
Objective: To study the subcellular localization of severe fever with thrombocytopenia syndrome virus (SFTSV) in macrophages and understand the replication and assembly mechanism of SFTSV in host cells.
Methods: Using two types of human macrophage cell lines THP-1 and U937, the study analyzed the intracellular colocalization of SFTSV with Golgi apparatus and endoplasmic reticulum by immunefluorescence staining and confocal microscopy.
Results: SFTSV infected macrophage cell lines THP-1 and U937.
Background: Severe fever with thrombocytopenia syndrome (SFTS) is an emerging infectious disease caused by the SFTS virus (SFTSV) with an average fatality rate of 12%. The clinical factors for death in SFTS patients remain unclear.
Methods: Clinical features and laboratory parameters were dynamically collected for 11 fatal and 48 non-fatal SFTS cases.
To understand the maintenance and transmission of SFTS virus, the potential vector ticks were collected from sheep, cattle and dogs in the endemic areas of SFTSV in Shandong Province. Among the collected ticks, the dominant species was H. longicornis ticks.
View Article and Find Full Text PDFSevere fever with thrombocytopenia syndrome bunyavirus is a newly emerging virus in China, enveloped with a tripartite, single-stranded RNA genome of negative polarity. The regulatory elements for viral transcription and replication, as well as encapsidation and packaging signals, are thought to be located within these noncoding regions (NCRs). The terminal nucleotides are genus specific and highly conserved.
View Article and Find Full Text PDFSevere fever with thrombocytopenia syndrome bunyavirus (SFTSV) is a novel phlebovirus, causing a life-threatening illness associated with the symptoms of severe fever and thrombocytopenia syndrome. The sequence and structure of the genome have already been illustrated in previous study. However, the characteristics and function of the structure and non-structure proteins is still unclear.
View Article and Find Full Text PDFZhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi
June 2011
Objective: To obtain recombinant human anti-EV71 antibodies from a EV71-associated hand-foot-and-mouth disease patient-derived antibody phage library.
Methods: A combinatorial human scFv library to enterovirus 71 (EV71) virus was constructed using antibody genes harvested from the blood of EV71 virus patients. The library was panned and selected by using purified VP1 protein of EV71 virus with phage display.
Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi
April 2011
Objective: To secreted express envelope glycoprotein (E) of dengue virus type 2 extracellularly.
Methods: The entire prM/E gene was amplified by RT-PCR. An optimized signal sequence gene from Japanese encephalits virus (JEV, SA14-14-2 strain) was introduced using fusion PCR.
Background: Heightened surveillance of acute febrile illness in China since 2009 has led to the identification of a severe fever with thrombocytopenia syndrome (SFTS) with an unknown cause. Infection with Anaplasma phagocytophilum has been suggested as a cause, but the pathogen has not been detected in most patients on laboratory testing.
Methods: We obtained blood samples from patients with the case definition of SFTS in six provinces in China.
Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi
June 2010
Objective: To observe the ability of dengue virus type 1-4 envelope domain III fusion protein to inhibit virus infection and analyze the neutralizing ability of polyclonal antibodies against rE III.
Methods: After being connected by linker peptide, E III protein of Dengue virus serotypes 1-4 were expressed in E coli BL21 (DE3) then purified. Fusion proteins were verified by Western Blot and ELISA.
Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi
February 2010
Objective: Development of pseudoviral competitive internal controls for RT-PCR laboratory detection of dengue virus.
Methods: The internal controls target gene were obtained by insertion of a 180 bp non-related DNA fragment into RT-PCR detection target of dengue virus between the forward and reverse PCR primer binding regions. A yellow florescence protein reporter gene was induced at downstream of internal controls target gene via internal ribosome entry site gene.
Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi
December 2009
Objective: In order to lay the groundwork for studying the novel vaccine Identified.
Methods: (1) Two replicons were constructed. One's prM/E gene was deleted completely (Full AprM/E Replicon), the other's prM/E gene was deleted partially (213 bp of C terminal of E gene was reserved; Partial delta prM/E Replicon), and the deleted parts was replaced as the MCS.
Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi
December 2009
Objective: To expression prM/E gene of dengue virus type I in mammalia cells.
Methods: The full-length prM/E gene of dengue virus type I strain GZ01/95 was amplified by RT-PCR, the signal peptide preceding the prM gene was added or the carboxyl-terminal 20% of DEN-1 E was replaced with the corresponding JE sequence in the meanwhile, and three of the constructions were cloned into the pcDNA5/FRT.Then they were transfected into 293T cells by lipofectamine respectively.
Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi
April 2009
Objective: To develop and evaluate a multiplex detection of IgM antibodies to pathogens caused viral hemorrhagic fever.
Methods: The nucleocapsid proteins (NP) of HTN, SEO, Puu MBV, Lassa, RFV and HPS viruses expressed in prokaryotic cells and purified were coupled to 7 different xMAP fluorescent microbeads. The assay was evaluated and optimized when screened against a panel of reference sera collected from HFRS patients, and compared to commonly used MacELISA Kits.
The prM/E gene of DV2 was cloned into the JEV (SA14-14-2 strain) replicon vector which had been constructed previously, and the resulting recombinant plasmid was named pPartialdeltaprM/E. The constructed chimeric clone was linearized and then was transcripted into RNA in vitro. The produced RNA was transfected into BHK-21 cells.
View Article and Find Full Text PDFZhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi
June 2008
Objective: To observe the ability of dengue virus recombinant envelope protein domain expressed in E. coli to inhibit virus infection and induce the neutralizing antibody.
Methods: E III protein of Dengue virus serotypes 1-4 were expressed in E.