2021年04期目次

  • Effect of bifidobacterium on mucosal immunity of bivalent oral poliomyelitis vaccine of types Ⅰ+ Ⅲ

    ZHAO Ting;SHI Hong-yuan;LI Jing;CHEN Shi-yi;ZHAO Yu-ping;YANG Xiao-lei;LI Guo-liang;YANG Jing-si;Institute of Medical Biology,Chinese Academy of Medical Sciences and Peking Union Medical College;

    Objective To investigate the effect of bifidobacterium on mucosal immunity induced by bivalent oral poliomyelitis vaccine(bOPV)of types Ⅰ+ Ⅲ. Methods Polio receptor transgenic mice were inoculated with two doses of b OPV at an interval of 28 d. The mice in test group were inoculated with bifidobacteria 7 d before each dose of vaccine by gavage,while those in control group were fed with PBS. Stool samples were collected from the mice on days 3 and 7 after each dose of vaccine,in which the levels of polio-specific IgA and virus shedding were measured. Cytokine secretion level and lymphocyte subsets in intestinal lymph nodes of mice were determined after vaccination. Results The levels of poliospecific mucosal IgA in mice in test group on day 3 after immunization with the first and the second doses of vaccine were significantly higher than those in control group(P < 0. 05 or P < 0. 01). The positive conversion rate of IgA of mice in test group on day 3 after immunization with the first dose of vaccine was significantly higher(P < 0. 05),while the virus shedding level of typeⅠwas significantly lower(P < 0. 01),than those in control group. The immunization with bOPV activated more B lymphocytes(P < 0. 000 1)and more Th1 pathway cytokines IL-1 and TNF-α(P < 0. 05)in mice in test group. Conclusion Application of bifidobacteria before immunization with bOPV increased the secretion level of poliospecific mucosal immune IgA,which showed a certain ability in slowing down the virus shedding.

    2021 04 v.34 [Abstract][OnlineView][Download 1074K]

  • Preparation,characteristics and immune effect of modular HBc-VLPs carrier microneedle vaccine

    LIU Di;GAO Hui-xian;YAN Ping-mei;LI Na;LI Bo;WU Xiao-ying;Department of Biology,Taiyuan Normal University;

    Objective To prepare modular HBc-VLPs carrier microneedle vaccine and analyze its characteristics and immune effect. Methods By directional transformation of hbc gene sequence,HBc(A80C)monomer protein was prepared,pu-rified and self-assembled to form HBc-VLP nanocarriers. The influenza virus antigen M2e peptide as model antigen was connected to Cys residue of HBc-VLPs by Sulfo-SMCC to prepare M2e-HBc-VLPs model antigen. Antigen particles were mixed in a rapid-dissolving material as a matrix to prepare a coated microneedle vaccine which was subjected to gel puncture,skin puncture and release tests. The antigenicity of M2e-HBc monomer was analyzed by Western blot. BALB/c mice were immunized with the coated microneedle vaccine by intradermal injection at dosages of 50 and 100 μg respectively,of which the serum specific antibody titer was determined by indirect ELISA,and the T lymphocyte level by flow cytometry. Results HBc(A80C) monomer was assembled to form HBc-VLPs spherical carrier at a particle size of about 30 nm. Gel and skin puncture tests showed that the antigen reached the epidermal layer,of which more than 90% was released within 15 min. The conjugated M2e antigen specifically bound to the corresponding antibody. The vaccine effectively stimulated mice to produce antigen-specific IgG antibodies and activated the cellular immunity,which validated the effectiveness of the model vaccine. Conclusion The HBc-VLPs carrier with site-directed coupling function can carry a variety of antigen modules to form a targeted vaccine,shorten the time required for vaccine preparation and evaluation,and accelerate the vaccine development process. Large-scale rapid vaccination may be performed through skin immunization with microneedles. The research results lay a foundation of application of HBc-VLPs as a modular vaccine carrier,which can promote the development of modular microneedle vaccines.

    2021 04 v.34 [Abstract][OnlineView][Download 1361K]

  • EEffect of interferon-induced transmembrane protein 2 on proliferation of encephalomyocarditis virus in vitro

    BI Ying-jie;XIE Jing-ying;XU Shu-juan;DENG Ying-ying;LI Zhuo;FENG Ruo-fei;Key Bio-engineering and Technology Laboratory,Biomedical Research Center of the Northwest Minzu University;

    Objective To investigate the effect of interferon-induced transmembrane protein 2(IFITM2)on proliferation of encephalomyocarditis virus(EMCV) in vitro. Methods HEK293 cells were transfected with recombinant plasmid pcDNA3. 1-IFITM2 and siRNA sequence targeting IFITM2 respectively,and determined for IFITM2 mRNA transcription level by qPCR and IFIMT2 protein expression level by Western blot. HEK293 cells were infected with 100 TCID50 of EMCV,based on which the effects of overexpression and inhibiting expression of IFITM2 on the virus proliferation were evaluated by determination of virus copy number and virus titer. The HEK293 cells in which IFITM2 was overexpressed were infected with EMCV at a MOI of 3 and evaluated for the effect of IFITM2 overexpression on adsorption and entry of EMCV by determination of virus copy number. Results Both the mRNA transcription and protein expression levels of IFITM2 in HEK293 cells transfected with recombinant plasmid pcDNA3. 1-IFITM2 increased significantly(P < 0. 001)while those transfected with siRNA sequence decreased significanthly(P < 0. 001). Both the copy number and titer of EMCV in HEK293 cells in which IFITM2 was overexpressed decreased significantly(P < 0. 05),while those in the cells in which IFITM2 expression was inhibited increased significantly(P < 0. 001). However,the number of virus particles adsorbed in HEK293 cells in which IFITM2 was overexpressed showed no significant change(P > 0. 05),while that of virus particles entered decreased significantly(P < 0. 01). Conclusion IFITM2 inhibited the proliferation of EMCV in HEK293 cells mainly at the early stage of viral replication.

    2021 04 v.34 [Abstract][OnlineView][Download 1204K]

  • Expression of KCNMA1 in pancreatic cancer cell lines and action mechanism of expressed product

    YAN Jiang-yu;ZHANG Ce;Key Laboratory of Cellular Physiology,Shanxi Medical University;

    Objective To express KCNMA1 in pancreatic cancer(PC) cell lines and investigate its role in the proliferation,apoptosis,migration and invasion of PC cells. Methods KCNMA1 was cultured in PC cell lines Patu-8988 T,MiaPaCa-2,PANC-1,Capan-1,AsPC-1,CFPAC-1 and BxPC-3 respectively,of which the expression was determined by RT-PCR. The KCNMA1 gene in PC cells was silenced by small interfering RNA(siRNA),while the cell proliferation level was determined by CCK-8 method,the apoptosis by flow cytometry,the expressions of apoptosis-related proteins by Western blot,and the migration and invasion by Transwell assay. Results The expression levels of KCNMA1 in MiaPaCa-2 and PANC-1 cells were significantly higher than those in other cell lines. Compared with that in normal control(NC) group,the A450 values of MiaPaCa-2 and PANC-1 cells 24 h after transfection with siRNA increased significantly(P < 0. 05),while the proportion of apoptotic cells increased to 12. 72% and 25. 1% respectively,the expressions of BAX,Cleaved-PARP1 and Actived-Caspase-3 protein increased,the numbers of migrated MiaPaCa-2 and PANC-1 cells decreased to 103. 8 ± 10. 6 and 117 ± 12. 2,and those of invasive cells to 63 ± 5. 8 and 116. 8 ± 9. 8,respectively(each P < 0. 01). Conclusion KCNMA1 is highly expressed in PC cell lines MiaPaCa-2 and PANC-1,while silencing KCNMA1 gene can effectively inhibit the proliferation,migration and invasion and promote the apoptosis of PC cells.

    2021 04 v.34 [Abstract][OnlineView][Download 1334K]

  • Prediction of structure and B cell epitope of M protein of SARS-CoV-2

    YE Xiao-xian;LI Ming-yang;ZHU Shan-li;CUI Huai-rui;WANG Yu;ZHANG Li-fang;Institute of Molecular Viruses and Immunology,Department of Anatomy,School of Basic Medicine Sciences,Renji College,Wenzhou Medical University;

    Objective To predict the structure and possible B cell dominant epitopes of M protein of SARS-CoV-2.Methods The amino acid sequence of SARS-CoV-2 M protein was searched from the NCBI GenBank database and analyzed by using computer-aided bioinformatics software. The physic-chemical properties, secondary structure and tertiary structure of M protein were predicted by using ProtParam,SOPMA,GOR IV and Swiss Model on ExPASy server respectively,while the transmembrane region by using TMHMM and Phobius online software. The comprehensive analysis of hydrophilicity,flexibility,surface possibility,antigenicity and polarity were performed by using TMHMM and Phobius online software in combination with DNASTAR software. The homology of Coronavirus M protein was analyzed by using Vector NTI software. Results The SARS-COV-2 M protein consisted of 222 amino acids,which was a stable transmembrane protein mainly composed of α helix and random coil. Comprehensive analysis of various parameters speculated that the possible B cell dominant epitope was amino acids 5-14(NGTITVEELK). The M protein of SARS-CoV-2 showed the highest homology of 90% to those of SARS-CoV coronavirus and Bat SARS-like CoV. Conclusion The peptides 5-14 was a possible B cell dominant epitope of SARS-CoV-2 M protein. This study provided a reference for the experimental determination of B cell epitope of SARS-CoV-2 M protein and immune recognition,and laid a foundation of development of vaccines.

    2021 04 v.34 [Abstract][OnlineView][Download 1528K]

  • Preparation and identification of rabbit serum antibody against recombinant cyclin-dependent kinase 4

    CI Xing-yuan;WANG Ai-li;YE Yi-ming;XU Si-shi;WANG Jia-bei;PAN Yi-qi;CHEN Jun;ZHANG Li-fang;ZHAO Kong-nan;Institute of Molecular and Immunology,Department of Medical Microbiology and Immunology,Wenzhou Medical University;

    Objective To prepare the rabbit serum antibody against recombinant cyclin-dependent kinase 4(CDK4).Methods CDK4 gene was designed,synthesized and cloned into vector pET21a(+). The constructed recombinant plasmid pET21a(+)/CDK4 was transformed to competent E. coli BL21(DE3) and induced with IPTG. The expressed product was purified by nickel ion chelate affinity chromatography,mixed with an equal volume of Freund adjuvant and injected s.c. into rabbits in several sites,every other week for 3 times. Serum samples were collected from the rabbits at weeks 0,2,4,6 and 8 after immunization and determined for antibody titer by indirect ELISA,and for antibody specificity by immunofluorescence assay(IFA)and immunohistochemical assay. Results Restriction analysis and sequencing proved that recombinant plasmid pET21a(+)/CDK4 was constructed correctly. The relative molecular mass of recombinant CDK4 protein was 19 100. The serum CDK4 antibody reached a titer of 1∶163 840 in rabbits at week 8 after immunization,which recognized the CDK4 protein expressed in Siha cells and cervical cancer tissue. Conclusion Rabbit serum CDK4 antibody with high titer and specificity was successfully prepared,which laid a foundation of further study on the biological characteristics of CDK4.

    2021 04 v.34 [Abstract][OnlineView][Download 1426K]

  • Determination of minimum detection limit of detection kit for nucleic acid of 2019 new coronavirus(fluorescence PCR)

    DONG Jiang-kai;HUANG Qing-hong;FAN Juan;HUANG Ying-yan;ZHANG Yun-tao;YANG Xiao-ming;Shanghai GeneoDx Biotech Co.,LTD.;

    Objective To determine the minimum detection limit of the detection kit for nucleic acid of 2019 new coronavirus(2019-nCoV)(fluorescent PCR),manufactured by Shanghai GeneoDx Biotech Co.,LTD.,and verify whether2019-nCoV at the minimum detection limit level may be detected by the kit. Methods By setting up five concentration gradients of 62 500,12 500,2 500,500 and 100 copies/mL for three repeat tests on 20 different clinically positive samples,the minimum detection limit range was preliminarily determined. Three consecutive batches of detection kit were used for determination of 20 different samples at five concentration gradients of 1 500,700,500,300 and 150 copies/mL for 20 times to further determine the minimum detection limit. Another 25 different clinical positive samples were diluted to the concentrations at minimum detection limit level and determined by three consecutive batches of kits for 20 times to verify minimum detection limit. Results All the positive detection rates of samples at concentrations of 62 500,12 500,2 500 and 500 copies/mL were 100%,while that at a concentration of 100 copies/mL was not more than 55%,and the minimum detection limit was preliminarily determined as 100～500 copies/mL. Twenty repeat tests showed that the positive detection rates of sample at concentrations of 1 500,700 and 500 copies/mL were not less than 95%,while those at 300 and 150 copies/mL were less than 95%,and the minimum detection limit was determined as 500 copies/mL.However,all the positive detection rates of 25 positive samples diluted to the concentration at minimum detection limit level were more than 95%. Conclusion The minimum detection limit of detection kit for nucleic acid of 2019-nCoV(fluorescent PCR)manufactured by Shanghai GeneoDx Biotech Co.,LTD. was determined as 500 copies/mL. It has been verified that the virus samples at the concentration of minimum detection limit could be detected by the kit,which meets the relevant national requirements for the minimum detection limit of detection reagents.

    2021 04 v.34 [Abstract][OnlineView][Download 1280K]

  • Molecular epidemiological characteristics of rotavirus VP7 gene in Zhengding County of Hebei Province,Xiangtan County of Hunan Province and Yuhuan City of Zhejiang Province,China from 2016 to 2017

    CHEN Hong;LI Qing-liang;DUAN Kai;SHI Chen;ZHANG Dong;DONG Ben;BAI Xuan;QIAO Jian;XU Ge-lin;YANG Xiao-ming;GAO Zhao;LI Fang-jun;LV Hua-kun;ZHOU Hai-song;YAN Ting-dong;SHI Hai-yun;Wuhan Institute of Biological Products Co.,Ltd.;

    Objective To analyze the molecular epidemiological characteristics of VP7 gene of the rotavirus(RV) in Zhengding County of Hebei Province,Xiangtan County of Hunan Province and Yuhuan City of Zhejiang Province,China from 2016 to 2017. Methods Stool samples of patients infected with RV in the three regions were collected from 2016 to2017,from which VP7 gene fragments were amplified by RT-PCR and subjected to sequencing and typing. The homology of nucleotide sequence of VP7 gene was analyzed,based on which a phylogenetic tree was constructed. The nucleotide sequences were translated to amino acid sequences,based on which the difference in amino acid sequences of prototype strain,UK-human reassortant vaccine virus strain and the isolates from the samples of the three regions were analyzed.Results The sequencing result showed four 4 G genotypes,G1,G2,G3 and G9,of which of proportions were 5. 1%(10/199),21. 6%(43/199),6. 0%(12/199)and 67. 3%(134/199)respectively. The RV of four G genotypes showed f ar evolutionary distances to the corresponding prototype strains,while the homologies of nucleotide sequences were90. 6% ～ 91. 9%,83. 5% ～ 94. 0%,82. 6% ～ 97. 2% and 75. 8% ～ 89. 8% respectively. However,compared with prototype strain and UK-human reassortant vaccine virus strain,there were difference at 15,12,17 and 32 amino acid sites of the four G genotypes respectively. Conclusion The dominant epidemic strains of RV of G genotype are different in different regions at the same time,and the dominant subtype of epidemic strain G9 changes over time,thus the epidemiological surveillance of the virus should be strengthened.

    2021 04 v.34 [Abstract][OnlineView][Download 2015K]

  • Expression of Mincle in human peripheral blood cells and its clinical significance

    LI Shu-ping;CHEN Yun;CUI Tian-jiao;WANG Xiao-hua;LI Jin-hong;ZHENG Zhi-hua;Department of Nephrology,The Seventh Hospital Affiliated to Zhongshan University;

    Objective To investigate the distribution of macrophage inducible C-type lection(Mincle) in human peripheral blood cells and analyze the correlation of its expression level to the clinical data. Methods Fluorescent antibodylabeled human peripheral blood cells were determined by flow cytometry,while the expression of Mincle in various peripheral blood cells by fluorescent signal combined with FSC/SSC. The correlation of Mincle expression level to clinical indexes was assessed by bivariate linear correlation analysis using SPSS 25 software. Results Mincle was expressed in CD11b-positive myeloid leukocytes but not in CD11b-negative ones. The expression level of Mincle was the highest in monocytes,followed by that in granulocytes. However,no Mincle expression was detected in lymphocytes. The expression level of Mincle in monocytes was correlated with the erythrocyte sedimentation rate(ESR). Conclusion Mincle was mainly expressed in monocytes and granulocytes but not in lymphocytes,of which the expression level was the highest in partial monocytes. The Mincle expression level in monocytes showed a certain correlation to the clinical data.

    2021 04 v.34 [Abstract][OnlineView][Download 1350K]

  • Meta-analysis of safety of vaccination with tetanus,diphtheria and pertussis combined vaccine use in pregnancy

    ZHANG Xue;ZUO Wei-lun;Institution of Medical Biology,Chinese of Medical Science & Peking Union Medical College,Yunnan Key Laboratory of Vaccine Research & Development on Sever Infections Diseases;

    Objective To analyze the safety of vaccination with tetanus,diphtheria and acellular pertussis combined vaccine(TdaP) use in pregnancy. Methods Databases including China National Knowledge Infrastruture(CNKI),China Biology Medicine Disc(CBM),WanFang Datebase(WF),PubMed,Embase and Cochrane Library were searched for randomized controlled trials(RCTs)on the safety of TdaP in pregnancy from the establishment of the databases to April 2020.Meanwhile,manual search was conducted,and references of included literature were retrieved. Meta-analysis was conducted by using RevMan 5. 3 software. Results A total of six published articles on RCT were included. Meta-analysis showed that,compared with those in control group,the combined RR values for incidences of pain,swelling,redness,fever,headache,fatigue,muscle aches,premature delivery,severe adverse reactions,congenital abnormalities and severe adverse reactions in newborn in trial group were 3. 74(95% CI :1. 43 ～ 9. 76),6. 69(95% CI:3. 92 ～ 12. 34),2. 20(95% CI 1. 60 ～ 3. 02),1. 55(95% CI:0. 45 ～ 5. 30),1. 09(95% CI:0. 84 ～ 1. 41),0. 93(95% CI:0. 52 ～ 1. 67),1. 11(95% CI:0. 20 ～ 6. 61),1. 48(95% CI:0. 74 ～ 2. 98),0. 98(95% CI:0. 71 ～ 1. 34),0. 74(95% CI:0. 37 ～1. 50)and 0. 83(95% CI:0. 51 ～ 1. 35)respectively. Conclusion The vaccination with TdaP in pregnancy does not increase the risk of other adverse reactions except for the slightly higher incidences of local pain,redness and swelling.

    2021 04 v.34 [Abstract][OnlineView][Download 1585K]

  • Prediction of relationship between high-mobility group-box transcription factor 1 and breast cancer by bioinformatics analysis

    WANG Jun-mei;ZHANG Yi-qiang;HUANG Yan;LI Xue-qing;PEI Jin-hong;Department of Biochemistry and Molecular Biology,Changzhi Medical College;

    Objective To analyze the relationship between high-mobility(HMG)-box transcription factor 1(HBP1) and breast cancer. Methods The mutation status of HBP1 gene in breast cancer,expression level of HBP1 gene in breast cancer and its relationship to survival period,and the co-expressed gene of HBP gene as well as its relationship to HBP1 gene in breast cancer were analyzed by bioinformatics analysis. Results The overall mutation rate of HBP1 in breast cancer was 0. 00%. The expression level of HBP1 protein in breast cancer was significantly lower than that in normal tissues(P < 0. 01),which was positively related to the prognosis of breast cancer(P < 0. 001). BTG1,CREBBP and SH3 BGRL were related molecules to the biological function of HBP1. In breast cancer,the expression level of HBP1 was positively related to thoses of BTG1 and CREBBP(P < 0. 01),while was negatively related to that of SH3 BGRL3(P < 0. 01).Conclusion The expression of HBP1 gene influenced the occurrence and development of breast cancer and was positively related to the prognosis of the disease.

    2021 04 v.34 [Abstract][OnlineView][Download 1497K]

  • Seroepidemiological survey of hepatitis B among population Heyuan City,Guangdong Province,China in 2018

    WANG Hai;HUANG Feng-guang;YANG Fen;YE Ling;WU Man-ju;CHEN Jun-hu;GONG Li-fen;Center for Disease Prevention and Control of Heyuan City;

    Objective To investigate seroepidemiology hepatitis B(HB)in Heyuan City,Guangdong Province,China,so as to understand the immunization level of target population and provide a scientific evidence for formulating immunization strategies and measures. Methods A total of 300 serum samples were collected from outpatients or persons receiving physical examination and determined for HB antibody level,based on which the positive rates of HB antibody in the populations of both genders and at various ages were analyzed. Results The positive rates of HBsAg,HBsAb and HBcAb in the 300 subjectes were 5. 33%,67. 33% and 44. 33%,respectively,which showed no significant difference in two genders(P > 0. 05). However,there were significant differences between the HBsAg positive rates of population at ages of 0 month ～ less than 5 years and 15 ～ less than 79 years,and between those at 5 ～ less than 15 years and 15 ～less than 79 years(each P < 0. 001). The HBsAb positive rate in the population at ages of 0 month ～ less than 5 years showed significant difference with that at 5 ～ less than 15 years(P < 0. 001),while the HBcAb positive rate in the population at ages of 5 ～ less than 15 years with that at 15 ～ less than 79 years(P < 0. 01). Conclusion Moderate prevalence of HB among the population in Heyuan City is observed,indicating that HB is still a major public health problem. There is a certain distance from the target task of controlling the HBsAg carrier rate of children at ages of less than 5 years below 0. 1% by 2030. The relevant population should be re-vaccinated with HB vaccine if necessary to reduce the immunization blank population.

    2021 04 v.34 [Abstract][OnlineView][Download 1436K]

  • Development,optimization and verification of a method for detection of residual β-propiolactone content in inactivated SARS-CoV-2 vaccine (Vero cells) by gas chromatography

    YANG An-na;WANG Yin;YANG Dong-sheng;ZHOU Yan-ping;TU Jing;LU Jia;LI Qian;SHI Jin-rong;WANG Ze-jun;SHEN Shuo;Wuhan Institute of Biological Products Co.,Ltd.;

    Objective To develop,optimize and verify a gas chromatography assay for determination of β-propiolactone(BPL)residue in inactivated SARS-CoV-2 vaccine(Vero cells). Methods The condition for gas chromatography was as follows: The inlet temperature was 180 ℃,while the shunt ratio was 2∶1,the carrier gas was nitrogen,the detector temperature was 250 ℃,and the operation time was 10 min. The column temperature condition(heating and isothermal procedures),flow rate(1 and 3 mL/min)and sample load(1 and 0. 2 μL)were optimized. The developed method was verified for specificity,limit of detection(LOD),limit of quantitation(LOQ),linear range,reproducibility and durability,and used for determination of change of BPL content during inactivation and hydrolysis in production procedure of inactivated SARS-CoV-2 vaccine(Vero cells). Results The optimal column temperature condition,flow rate and sample load were isothermal procedure,3 mL/min and 0. 2 μL respectively. The retention time of BPL peak of mixture of BPL reference and bulk of vaccine was 2. 328 min,which was consistent with that of BPL reference. All the RSDs of peak area and retention time of BPL reference in six consecutive sample loadings and those at various detector and inlet temperatuers were not more than 4. 0%. The peak area of BPL reference showed good linear relationship to the concentration at a range of 3. 125 ～ 500 μg/mL,of which the equation was y = 1. 399 1 x-0. 558 6(r = 0. 999 9).The LOQ and LOD of the developed method were 2. 530 and 0. 843 μg/mL respectively. The BPL content during inactivation was higher than the LOQ,while that after hydrolysis was remarkably lower than the LOD. No BPL was detected in the bulk of vaccine. Conclusion A gas chromatography assay for determination of BPL residue in inactivated SARS-CoV-2 vaccine(Vero cells)was developed,which showed high specificity,precision and durability,and might be used for monitoring the BPL content in various steps of vaccine production procedure.

    2021 04 v.34 [Abstract][OnlineView][Download 1491K]

  • Interlaboratory verification of fluorescent quantitative PCR for detection of three extraneous avian viruses in influenza vaccine virus strains

    WANG Shu-jing;FU Rui;QIN Xiao;LIU Chao-yang;HU Ya-ling;WANG Zeng;HAN Bin;WANG Kui;YUE Bing-fei;National Institute for Food and Drug Control;

    Objective To verify the fluorescent quantitative PCR(Q-PCR)for detection of three extraneous avian viruses in influenza vaccine virus strains among various laboratories. Methods The Q-PCR method for detection of avian adenovirus typesⅠ(chicken embryo lethal orphan virus,CELO)and Ⅲ(egg drop syndrome virus,EDS)as well as avian leukemia virus(ALV) in influenza vaccine virus strains were subjected to interlaboratory verification for sensitivity,specificity,reproducibility,virus contamination mimic experiments and blind sample detection by four laboratories.Results The verification results showed that the R~2 values of standard curves of Q-PCR methods by various laboratories were more than 0. 99,while the amplification efficiencies were 93. 263% ～ 103. 105%,and the sensitivities reached 1×10~1 copies/μL. Only the positive target viruses were detected,while no amplification curves of other viruses appeared.All the coefficients of variation(CVs)of the cycle thresholds and copies in intra-and inter-assays were less than 15%. The minimum detection limits of CELO,EDS and ALV in virus contamination experiments were 1 × 10~(-7),1 × 10~(-4) and 1 ×10~(-3) virus incorporation respectively,and the detection results of blind samples by four laboratories were in agreement.Conclusion The verification results of Q-PCR method for detection of extraneous avian viruses by various laboratories were in agreement. The method showed high sensitivity,specificity and reproducibility,which might be used for the rapid detection of extraneous avian virus in influenza vaccine virus strains.

    2021 04 v.34 [Abstract][OnlineView][Download 1513K]

  • Development verification and application of indirect ELISA for pyolysin antibody against Trueperella pyogenes in goats

    XU Deng-feng;ZHANG Su-hui;YU Yuan-di;XU Guo-yang;ZHAO Yang-yang;FU Li-zhi;SHEN Ke-fei;Chongqing Academy of Animal Sciences;

    Objective To develop verify and apply an indirect ELISA for pyolysin antibody against Trueperella pyogenes in goats. Methods Recombinant pyoysin(r PLO) was expressed in E. coli and purified by Ni-NTA chromatography. An indirect ELISA was established for detection of anti-PLO IgG antibody against T. pyogenes in goats was developed using r PLO as coating antigen and HRP-labeled rabbit anti-goat IgG as the secondary antibody. The concentration of coating antigen(3. 5,0. 7,0. 14 and 0. 05 μg/mL)and the dilution of secondary antibody(1∶10 000,1∶20 000 and 1∶40 000)were optimized by block titration. The developed method was verified for specificity,sensitivity and reproducibility.Fifteen goat serum samples naturally infected with T. pyogenes were determined by agarose gel diffusion test and the optimized indirect ELISA respectively,and the coincidence rate of results was calculated. A total of 360 clinical serum samples of goats were determined by the optimized indirect ELISA. Results The purified r PLO,with a relative molecular mass of about 34 000,reached a purity of 90%. The optimal concentration of coating antigen was 0. 14 μg/mL,while the optimal dilution of secondary antibody was 1∶40 000. The developed ELISA showed no cross reaction with the sera positive for E. coli Streptococcus,or Corynebacterium pseudotuberculosis. The positive serum samples at a dilution of 1∶800 was detected by the developed method. Both the coefficients of variation of determination result by the developed ELISA in the intra-batch and inter-batch repetitive tests were less than 10%. Both the positive rates of fifteen goat serum samples naturally infected with T. pyogenes determined by two methods were 100%,of which the incidence rate was 100%. However,the positive rate of 360 goat serum samples determined by the optimized ELISA ws 24. 1%(87/360).Conclusion The indirect ELISA for PLO antibody against T. pyogenes in goats was successfully developed,which showed high specificity,sensitivity and precision.

    2021 04 v.34 [Abstract][OnlineView][Download 1557K]

  • New technology for antigen design and its application in vaccine

    LI Xing-hang;YANG Xiao-ming;National Research Center of Engineering Technique for Combine Vaccine;

    Vaccinology is originated with Jenner and Pasteur who mimicked the protective effect of natural infection in humans by inoculation with inactivated or attenuated pathogens. This approach led to the vaccine development lasting for a century and effective prevention of various infectious diseases. However,effective vaccines against partial diseases have not been developed by the traditional method. With the development of various disciplines,new technologies and strategies such as structural vaccinology and vaccinomics have emerged in the field of vaccine design. These new technology platforms provide new ideas for designing antigens in new vaccines. This paper reviews the new technology for antigen design as well as its application and development in vaccines.

    2021 04 v.34 [Abstract][OnlineView][Download 1689K]

  • Progress in research on multivalent influenza vaccine and universal influenza vaccine

    XIE Qing-xin;LIU Yan-hong;LU Wei-dong;College of Pharmacy,Kunming Medical University,Yunnan Provincial Key Laboratory of Pharmacology of Natural Drugs;

    Vaccination is an important measure to prevent and control influenza. Common influenza vaccine includes trivalent and tetravalent vaccines. With the development of research on influenza vaccine,the current multivalent vaccines are optimized in aspects of the selection of vaccine virus strain,addition of adjuvant and improvement of production process. Due to the poor cross-protection of influenza vaccine among different subtypes,universal influenza vaccine which can induce broad spectrum and long-term immunity is the main trend of influenza vaccine development. In recent years,the discovery of antigens that can cause a wide range of viral responses,the modification of conserved region of the virus and the study on cellular immune mechanisms have provided new ideas for the development of universal influenza vaccine. In this paper,the current research status of influenza vaccine and the progress in development of universal influenza vaccine are reviewed.

    2021 04 v.34 [Abstract][OnlineView][Download 1515K]

  • Progress in research on antiviral mechanism of types Ⅰ and Ⅲ interferon

    JIN Li;SU Qiang;ZHOU Jian-hua;MA Zhong-ren;Center for Biomedical Research,Northwest Minzu University;

    Interferon(IFN)is an important antiviral cytokine which directly involves in the antiviral immune response of epithelium and mediates the induction of interferon stimulated gene(ISG) by innate natural immune response thus establishes the natural immune defense against virus infection rapidly. It is currently believed that types Ⅰ and Ⅲ IFN have redundant functions,such as antiviral,anti-tumor proliferation and immunomodulatory effects. Meanwhile,the signaling pathways mediated by the two types are similar to a large extent. Type Ⅰ interferon exerts antiviral effect in the early stage of viral infection as well as long-term immune function,which is rapid in response and strongly induces the production of ISGs. However,typeⅢ interferon is relatively slow in response and limit in induction of ISGs,which plays an important role primarily in mucosal immunity in innate immune response. In addition,gene expression results in the difference in evolution of IFN system,reflecting the diversity between species. In this paper,the progress in research on the roles of typesⅠand Ⅲ interferon in immunomodulation and antiviral fields in order to provide a reference for study on IFN-mediated immunity.

    2021 04 v.34 [Abstract][OnlineView][Download 1540K]

  • Progress in research on prophylactic mRNA vaccines

    LIU Jing-jing;LI Yu-hua;Department of Arbovirus Vaccines,National Institutes for Food and Drug Control;

    As a novel nucleic acid vaccine,mRNA vaccine has become a promising vaccine approach because of its safety, effectiveness, rapid production and low-cost, especially its unique advantage in prevention and control of emerging and sudden infectious diseases. With overcoming the issue of the instability and inefficient delivery,several mRNA vaccine platforms have been built and used for the prevention and control of infectious diseases as well as therapy of cancers. In this review we summarized the mRNA design,RNA delivery and research status of clinical or pre-clinical trials to expound the prospects in the field of preventing and controlling the infectious disease in the future.

    2021 04 v.34 [Abstract][OnlineView][Download 1629K]

  • Progress in research on human adenovirus vaccine

    XI Hua-long;GU Han-xue;SUN Bo;GU Tie-jun;Changchun BCHT Biotechnology Co.;

    Human adenovirus(HAdV)is a double-stranded DNA virus without envelope,which causes various diseases through different routes. At present,there are no effective drugs against HAdV. Commercial adenovirus vaccine is currently available only in the United States. However,adenovirus vaccine has not been approved in China,which is still in the early stage of research. In this paper,the development and application of HAdV vaccine are reviewed.

    2021 04 v.34 [Abstract][OnlineView][Download 1563K]


@2012 Editorial Department of "Chinese Journal of Biologicals"