关键词： 脊髓灰质炎疫苗   疑似预防接种异常反应   免疫策略   转换  

摘要： 目的比较辽宁省脊髓灰质炎(脊灰)疫苗免疫策略转换前后不同脊灰疫苗疑似预防接种异常反应(AEFI)发生率。方法通过中国AEFI监测系统收集辽宁省脊灰疫苗免疫策略转换前(2013年5月-2016年4月)三价脊灰减毒活疫苗(tOPV)和转换后(2016年5月-2019年12月)二价脊灰减毒活疫苗(bOPV)、脊灰灭活疫苗(IPV)AEFI数据,比较AEFI报告发生率。结果辽宁省免疫策略调整前tOPV和调整后bOPV和IPV的AEFI报告发生率(/10万)分别为6.03、11.11、27.80,其中一般反应分别为5.44、10.14、26.16,异常反应分别为0.28、0.62、1.29。在tOPV、bOPV、IPV异常反应中,过敏性皮疹报告发生率(/10万)分别为0.10、0.47、1.17,疫苗相关麻痹型脊灰报告发生率(/10万)分别为0.07、0.04、0.00。结论辽宁省脊灰疫苗免疫策略调整前后三种脊灰疫苗不良反应发生率在预期范围内,严重反应极为罕见。

关键词： Vero细胞   Ⅲ型脊髓灰质炎病毒   感染性滴度   D抗原  

摘要： 型脊髓灰质炎病毒经下游灭活工艺后抗原损失量较大,可以通过优化上游培养工艺来提高病毒产量。为研究病毒接种时间、感染复数(MOI)和病毒收获时间对Ⅲ型脊髓灰质炎病毒感染性滴度(CCID50)和D抗原的影响,优化Ⅲ型脊髓灰质炎病毒在Vero细胞中的增殖条件,为生产及毒种制备提供指导依据,本研究以接种胞龄和MOI为病毒接种的两个变量条件,以病毒滴度和D抗原含量为病毒收获液中病毒产量的判定指标。先固定MOI,在不同胞龄的Vero细胞上以相同MOI接种Ⅲ型脊髓灰质炎病毒,根据病毒收获液中病毒产量的结果筛选出产毒量最高的接种胞龄范围。然后以最佳产毒胞龄为固定条件,暨Vero细胞生长至96h^144h,以不同MOI接种病毒,进一步筛选出产毒量最高的MOI范围。结果显示,当MOI=0.003时,Vero细胞培养48h、72h、96h、120h和144h种毒组均在种毒后70h完全病变,Vero细胞培养120h种毒所得收获液D抗原含量和病毒滴度均为最高,分别为(331±31)DU/mL、(8.04±0.14)lgCCID50/mL。接种胞龄固定在96h^144h范围,当MOI为0.001、0.01和0.1时,细胞完全病变时间分别为81h、66h和55h,所得病毒收获液的D抗原含量分别为(301.1±13.8)DU/mL、(303.7±15.6)DU/mL和(272.6±19.6)DU/mL,滴度分别为(7.88±0.13)lgCCID50/mL、(7.58±0.14)lgCCID50/mL和(7.75±0.13)lgCCID50/mL。本研究提示,相同MOI下,不同细胞胞龄种毒后细胞达到完全病变时间相同;细胞培养至平台期前后接种Ⅲ型脊髓灰质炎病毒所获得的收获液D抗原含量和感染性滴度均最高;MOI影响Ⅲ型病毒病变进程,但对病毒产量影响不大;90%以上Vero细胞发生病变、脱落时为最佳收获病毒时间点。

关键词： 脊髓灰质炎   实验室质量控制标准毒株   细胞敏感性  

摘要： 目的制备和滴定2019年河南省脊髓灰质炎病毒实验室质量控制标准毒株(Laboratory Quality Control Strain,LQCS)。方法中国Sabin参考毒株(China Sabin Test Reference Strain,CSTRS)扩增制备LQCS,以CSTRS为平行实验,进行三次独立实验完成LQCS滴定。结果LQCS RDⅠ型为108.48 CCID50/0.1 mL、RDⅢ型为108.57 CID50/0.1 mL、L20BⅠ型为107.82CCID50/0.1 mL和L20BⅢ型为107.20 CCID50/0.1 mL。结论河南省已制备可用于细胞敏感性实验的LQCS。

关键词： Sabin株脊髓灰质炎病毒   凝胶过滤层析   Capto   Core   400   复合型层析介质   参数优化  

摘要： 目的:分别用新型复合型层析介质Capto Core 400和传统介质Sepharose 6FF纯化Sabin株脊髓灰质炎病毒(Sabin strain poliovirus,sPV)去除宿主细胞蛋白(hostcell protein,HCP)和DNA,并且比较纯化结果及工艺参数。方法:用Capto Core 400与Sepharose 6FF纯化Vero细胞培养的Ⅰ、Ⅱ、Ⅲ型sPV浓缩液。分别检测纯化后sPV的D抗原含量、HCP残留量和DNA残留量,计算D抗原回收率、HCP和DNA去除率,并用t检验分析差异。结果:Capto Core 400与Sepharose 6FF纯化Ⅰ、Ⅱ、Ⅲ型sPV的D抗原回收率差异均无统计学意义(t值分别为1.09、1.08、1.02,P值均>0.05),但前者的Vero细胞HCP(t值分别为3.15、3.23、3.54)和DNA去除率均高于后者(t值分别为3.41、3.25、3.62),且差异有统计学意义(P值均<0.05)。Capto Core 400与Sepharose 6FF的介质使用体积比值为0.05,上样后纯化时间比值为0.04,工作缓冲液使用量比值为0.25,工艺线性流速比值为10,每升介质上样量比值为20,最大耐压力比值为2。结论:对比Sepharose 6FF,Capto Core 400可以更有效地去除Vero细胞HCP与DNA,各个工艺参数得到了优化,提高了sPV层析纯化的工作效率,并节约了时间和成本。

关键词： continuous quench-flow reactor   disinfection kinetics   EMA-qPCR   ozone disinfection   viral infectivity  

摘要： Aims A continuous quench-flow (CQF) reactor was developed to collect samples at the reaction times of less than one second. The reactor is applied to determine ozone disinfection kinetics of poliovirus and to study whether EMA-qPCR can assess the viral infectivity after ozone disinfection. Methods Ozone disinfection of poliovirus was conducted in the developed CQF, and the disinfection kinetics were tested in the range of 0 center dot 7-5 center dot 0 s at ozone concentration of 0 center dot 08 and 0 center dot 25 mg l(-1). Inactivation, damage on viral genome and damage on capsid integrity were determined by plaque assay, quantitative reverse transcription polymerase chain reaction (RT-qPCR) and ethidium monoazide treatment coupled with RT-qPCR (EMA-qPCR), respectively. Results By using CQF, 2 center dot 18 and 2 center dot 76 log(10)reductions were observed at the reaction time of 0 center dot 7 s and ozone concentration of 0 center dot 08 and 0 center dot 25 mg l(-1), respectively, followed by tailing. Ozone disinfection kinetics of poliovirus 1 were better fit by the efficiency factor Hom model than by the Chick-Watson model, or the modified Chick-Watson model. Kinetics observed were similar between RT-qPCR and EMA-qPCR assays at the reaction times of 5 s, viral concentration evaluated by EMA-qPCR was reduced in comparison to stable RT-qPCR results. Both assays still underestimated the virus inactivation. Conclusion The simple developed reactor can be used to investigate viral ozone disinfection kinetics and to elucidate inactivation characteristics or mechanisms at very short exposure times. Significance and Impact of the Study The developed CQF reactor is beneficial for better understanding of virus inactivation by ozone, and the reactor can be used to better elucidate disinfection kinetics and mechanisms for future research. This work constitutes an important contribution to the existing knowledge of the application and limitation of the EMA/PMA-qPCR to asses

关键词： Enterovirus   Environmental surveillance   Phylogenetic characterization   Epidemiology  

摘要： We present the results of environmental surveillance for poliovirus (PV) and non-poliovirus (NPEV) around the switch from trivalent to bivalent oral polio-vaccine (OPV) which occurred in China in May 2016. Sewage samples were collected in Jinan and Linyi city from 2015 to 2017. Enterovirus (EV) isolation, VP1 amplification, Sanger sequencing, and phylogenetic analyses were performed. Among105 sewage samples (36 in Jinan and 69 in Linyi), 101 were positive for EV, with 74.3% (78/105) PV-positive samples and 90.5% (95/105) NPEV-positive samples. A total of 893 EV isolates were obtained, including 326 (36.5%) PVs and 567 (63.5%) NPEVs. Echovirus (E) -11 was the most common serotype out of 18 detected NPEV types (120/567), followed by E-3 (75/567) and E-6 (74/567). PV2 vanished and PV3 came to be the ascendant PV type in sewage after May 2016. Eight PV isolates were judged as pre-vaccine-derived poliovirus (pre-VDPV) and no VDPV or wild PV isolates were monitored. Bayesian phylogenetics demonstrated global E-11 originated in 1876 and evolved with the estimated rate of 4.63 x 10(-3) nucleotide substitutions per site per year (s/s/y). Multiple circulating clusters that originated at different times were coexisting in Shandong province. The most recently common ancestor of global coxsackievirus B5 could date back to 1867, at the evolutionary rate of 3.95 x 10(-3) s/s/y. In conclusion, our study described the changes of PVs and NPEVs around the polio vaccine switch period and provided meaningful global molecular epidemiological data for further studies of EV-related diseases among the population.

关键词： HIV origins   The WetNet   experimentality   Africa   zoonosis  

摘要： The author analyzes the aftermath of Edward Hooper's suggestion that the trial of an oral polio vaccine (OPV) in the Belgian colonies of Africa engendered the pandemic form of the AIDS virus, HIV-1. In response to Hooper's book, The River(1999), the Royal Society in London held a conference to debate the origins of HIV. Examination of the quick dismissal of the OPV theory opens a space for legitimately challenging the widely held belief that the vaccine contamination question was convincingly resolved. This article interrogates the relationship between historiography and the making of scientific facts and history, suggesting that historians have been too credulous of scientists' testimony. The further result of the lack of a thorough analysis of the evidence backing the OPV hypothesis has resulted in a missed opportunity to readThe Riveras one of the few detailed accounts of the immense social, political, technological, and interspecies infrastructure constituted by Cold War vaccine production. This biomedical infrastructure dramatically changed the geographic and interspecies mobility of viruses in ways that may be impossible to reconstruct. Yet these potential transmission routes remain crucial to acknowledge. The COVID-19 pandemic draws attention to the critical importance of studying The WetNet, a concept coined by the author to name the conceptual and material infrastructures of inter- and intraspecies fluid bonding.

关键词： GAPIII   poliovirus   PVR knockout   unintentional propagation  

摘要： After eradication and containment of wild poliovirus (PV) and cessation of oral polio vaccinations, it is critical to minimize the risk of reintroducing PV into PV-free communities via facilities that handle the virus. The potential risk of unintentional PV propagation through unidentified contaminated materials is a serious issue. This study reports the generation of HeLa and RD-A cells deficient in functional CD155 gene ( increment PVR cells);these cells are not susceptible to PV but remain susceptible to other picornaviruses. These increment PVR cells will minimize the risk of unintentional transmission of PV and support performing the experiments more safely.

关键词： Vaccines   Infection   Diseases   SARS-CoV-2  

摘要： The back story Medical researcher Albert Sabin developed the conventional polio vaccine in the 1950s and 1960s, by growing the virus in non-human primates and cell cultures, until it adapted to those environments and was no longer good at infecting humans. Andino started working on the new vaccine in 2011, with colleagues including Andrew Macadam at the UK National Institute for Biological Standards and Control and others at the US Centers for Disease Control and Prevention. In 2015, the Bill and Melinda Gates Foundation in Seattle, Washington, agreed to fund a $150-million programme of simultaneous clinical trials and manufacturing of the new vaccine; the non-profit global health organization PATH, also based in Seattle, is coordinating the project.