关键词： 脊髓灰质炎病毒   型内鉴定   实时荧光定量逆转录-聚合酶链反应  

摘要： 目的评价脊髓灰质炎病毒(Poliovirus,PV)型内鉴定(Intratypic differentiation,ITD)的实时荧光定量逆转录-聚合酶链反应(Real-time reverse transcription-polymerase chain reaction,rRT-PCR)检测方法。方法对2013-2018年新疆环境污水中PV分离株,采用rRT-PCR方法进行PV ITD检测,分析5种版本rRT-PCR的疫苗衍生脊灰病毒(Vaccine derived poliovirus,VDPV)筛查假阳性率。结果 2013-2018年共分离到PV 1 792株,经rRT-PCR进行PV ITD检测,其中Ⅰ、Ⅱ、Ⅲ型PV分别为490株、372株、930株。从3.0版到5.0版rRT-PCR,PV ITD检测时间从160min缩短至137min,Ⅰ型、Ⅲ型VDPV筛查假阳性率分别从55.9%、41.1%降低至0.0%、0.2%;各版本rRT-PCR的Ⅱ型VDPV筛查假阳性率在44.0%-62.1%之间。结论 rRT-PCR方法的不断优化提高了PV ITD的检测准确性和及时性。

关键词： 二价脊髓灰质炎减毒活疫苗   损耗系数   影响因素  

摘要： 目的分析武汉市洪山区二价脊髓灰质炎减毒活疫苗(bOPV)损耗系数,了解其损耗原因及影响因素,为科学制定疫苗使用计划,规范疫苗管理提供依据。方法利用湖北省免疫规划信息系统及疫苗出入库管理系统等数据,设计专用调查表了解洪山区各预防接种单位bOPV接种及使用情况,采用描述性统计分析方法计算损耗系数。结果洪山区25家预防接种单位bOPV损耗系数为1.69。区卫计委直属门诊(1.35)和大型医院承办社区门诊(2.01)损耗系数之间的差异有统计学意义;bOPV接种周期1次/周(1.57)和2~6次/周(1.94)损耗系数之间的差异有统计学意义;疫苗启用废弃时间按<4 h(1.86)和>4 h(1.61)损耗系数之间的差异有统计学意义。做损耗系数相关因素的多因素回归分析发现,不同的门诊属性比接种周期和接种台数量对损耗因素的影响更大。2016和2017年损耗系数分别为2.09和1.75,经比较,两者差异有统计学意义。结论洪山区bOPV管理及使用比较规范,可通过加强监管考核、设置集中接种、规范宣传培训等手段进一步减少疫苗损耗。

关键词： 脊髓灰质炎病毒   脊髓灰质炎病毒疫苗,口服   龙胆紫   方法  

摘要： 目的 建立测定脊髓灰质炎病毒(poliovirus,PV)滴度的结晶紫染色法,并对该法进行验证。方法 基于PV的半数细胞培养感染量(50% cell culture infective dose,CCID50),用结晶紫染色和酶标仪确定PV的致细胞病变效应(cytopathic effect,CPE),并验证该法的准确度、精密度、线性范围和耐用性。分别采用建立的结晶紫染色法和传统的显微镜观察法测定66批口服Ⅰ型Ⅲ型脊髓灰质炎减毒活疫苗的病毒滴度,通过Pearson相关分析评价2种检测方法的相关性。将细胞板浸泡液盲传3代,观察培养物的CPE,并通过逆转录PCR确定病毒灭活情况。结果 用建立的方法测定理论滴度8.00、6.00和4.00 lgCCID50/ml的PV样品,回收率为98%~104%。细胞板内和板间测定结果的变异系数(coefficient of variation,CV)为0%~7.85%,同一工作日和不同工作日测定结果的CV分别不超过6.44%和5.27%,不同实验人员测定结果的CV不超过5.10%。该法测定PV滴度的线性范围为2.00~8.00 lgCCID50/ml。该法具有较好的耐用性,以不同细胞浓度(F=1.624,P=0.215)或不同培养时间(F=2.731,P=0.055)培养对PV滴度的测定结果没有影响。该法与显微镜观察法的检测结果具有相关性,Pearson相关系数0.918。实验用细胞板的浸泡液在Hep-2细胞上盲传3代后,培养物未观察到CPE,逆转录PCR测定结果为阴性。结论 建立的结晶紫染色法具有较好的安全性、准确度、精密度和耐用性,适用于口服脊髓灰质炎减毒活疫苗的病毒滴度测定。

关键词： 脊髓灰质炎后综合征   康复治疗   疗效  

摘要： 对2例脊髓灰质炎后综合征患者进行个体化的康复治疗,结果显示他们的疼痛明显减轻,运动功能明显提高。

关键词： 脊髓灰质炎病毒   Sabin株   C抗原   D抗原   分离  

摘要： 目的分离脊髓灰质炎病毒Ⅲ型Sabin株(sPVⅢ)C抗原和D抗原,并分析其部分特性。方法采用CsCl密度梯度离心法分离sPVⅢ的C抗原和D抗原,电镜观察病毒形态;SDS-PAGE检测病毒结构蛋白组成;微量BCA法检测蛋白浓度,计算每种抗原所占比例;细胞病变法检测病毒滴度。同时检测上样浓度及超离次数对C抗原纯度的影响。结果sPVⅢC抗原和D抗原在透射电镜下均呈球形,C抗原病毒颗粒密度较小,外壳大多较为松散,D抗原病毒颗粒密度较大,结构更稳固;C抗原由VP0、VP1、VP3蛋白组成,D抗原由VP1、VP2、VP3和VP4蛋白组成;C抗原和D抗原分别约占总蛋白含量的10%和90%,部分C抗原在分离前及分离过程中易产生聚集;D抗原的病毒滴度高于C抗原。降低上样浓度及进行二次超离可提高C抗原纯度。结论采用CsCl密度梯度离心法成功分离了sPVⅢ的C抗原和D抗原,C抗原滴度远低于D抗原,且部分易产生聚集,较难达到较高纯度。

关键词： 关节置换术,置换,髋   髋关节发育不良   脊髓灰质炎后综合征  

摘要： 目的探讨全髋关节置换术(THA)治疗脊髓灰质炎后遗症患者非瘫痪侧髋关节发育不良(DDH)的临床效果。方法回顾性分析2015年1月—2018年4月解放军东部战区总医院骨科收治的12例非瘫痪侧因DDH行THA的脊髓灰质炎后遗症患者的临床资料。其中男5例,女7例;年龄35~52岁,平均40.2岁;CroweⅠ型6髋,CroweⅡ型5髋,CroweⅢ型1髋。患者均采用后外侧入路治疗。观察患者手术时间、术后住院时间、出血量及并发症;对比手术前后患者Harris髋关节评分和疼痛VAS变化,采用Harris髋关节评分评定疗效。结果本组12例患者均顺利完成手术。手术时间51~91(64.9±11.0)min;术中出血量203~347(282.7±44.4)mL,术后急性失血量353~485(414.2±51.6)mL;术后住院时间4~9 d。术后无感染、骨折、神经血管损伤等并发症发生,手术切口均获得一期愈合。术后患者均获随访6~24个月,平均18个月。随访期间,患者下地行走良好,无假体脱位及假体松动发生。末次随访髋关节Harris评分较术前明显增加,分别为(62.8±7.52)和(90.2±2.64)分;末次随访VAS较术前明显降低,分别为0.00(0.00,0.00)分和4.00(3.00,6.00)分,差异均有统计学意义(t=11.913、Z=10.864,P值均<0.05)。术后髋关节Harris评分评定:优10髋,良2髋。结论通过对髋关节假体安放位置进行合理设计,采用THA治疗脊髓灰质炎后遗症患者非瘫痪侧DDH可以取得满意的疗效。

关键词： Polio   Vaccine-derived   iVDPV   Neutralization  

摘要： The use of the oral poliovirus vaccine (OPV) in developing countries has reduced the incidence of poliomyelitis by >99% since 1988 and is the primary tool for global polio eradication. Spontaneous reversions of the vaccine virus to a neurovirulent form can impede this effort. In persons with primary B-cell immunodeficiencies, exposure to OPV can result in chronic infection, mutation, and excretion of immunodeficiency-associated vaccine-derived polioviruses, (iVDPVs). These iVDPVs may have the potential for transmission in a susceptible population and cause paralysis. The extent to which sera from OPV recipients are able to neutralize iVDPVs with varying degrees of antigenic site substitutions is investigated here. We tested sera from a population immunized with a combination vaccine schedule (both OPV and inactivated polio vaccine) against a panel of iVDPVs and found that increases in amino acid substitution in the P1 capsid protein resulted in a decrease in the neutralizing capacity of the sera. This study underscores the importance of maintaining high vaccine coverage in areas of OPV use as well as active surveillance of those known to be immunocompromised. Published by Elsevier Ltd.

关键词： Immunoinformatics   multi-epitope   poliovirus   vaccine   molecular dynamic simulation  

摘要： Current vaccines used for polio are live attenuated oral polio vaccine and inactivated polio vaccine. Recently, in India, the relapse of virulence was observed in attenuated viruses resulting in catastrophic effects. Therefore, the need for the development of multi-epitope subunit vaccine was realised and an immunoinformatics approach to design a multi-epitope subunit vaccine was conceived. Capsid proteins of all the three types of polio strains were utilised to predict major histocompatibility complex class-1 as well as class-2 epitopes. The subunit vaccine was designed with beta-defensin at N-terminal followed by cytotoxic T-lymphocytes epitopes and helper T-lymphocytes epitopes connected by compatible linkers. The vaccine construct was further modelled and docked against TLR4 receptor. The high affinity of the construct towards the receptor was observed in the docking study and also substantiated by a 20 ns simulation of the complex. The vaccine construct was cloned in-silico for expression of the protein effectively in a prokaryotic system (Escherichia coli strain K12). Immuno-simulation of the construct was found to elicit immunoglobulin production effectively in the human body. This designed multi-epitope subunit vaccine is capable of immune response and further studies will help us understand the feasibility of this multi-epitope subunit vaccine.

关键词： Cuba   fractional dose   IPV   polio  

摘要： Backgrounds. Intradermal (id) fractional inactivated poliovirus vaccine ([fIPV] one fifth of normal IPV dose) is safe and immunogenic;however, id administration is perceived as difficult. We compared fIPV immunogenicity administered id or intramuscularly (im). Methods. This noninferiority trial was conducted among polio vaccine-naive Cuban infants who received 2 IPV doses at 4 and 8 months of age. Infants were randomized into 4 arms: (A) fIPV, 0.1 mL im;(B) fIPV, 0.2 mL im;(C) fIPV, 0.1mL id;and (D) IPV, 0.5 mL im. Blood collected before and after vaccinations was tested for poliovirus-neutralizing antibodies. Results. A total of 196 of 214 (91.6%) enrolled children completed study. Seroconversion after 2 IPV doses in each arm were as follows: (A) 97.3% (90.6-99.7), 98.7% (92.7-99.9), and 90.5% (81.5-96.1) for serotypes 1, 2, and 3, respectively;(B) 97.2% (90.3-99.7), 100%, 95.8% (88.3-99.1) for serotypes 1, 2, and 3, respectively;(C) 89.3% (71.8-97.7), 92.9% (76.5-99.1), 82.1% (63.1-93.9) for serotypes 1, 2, and 3, respectively;and (D) 100%, 100%, 100% for serotypes 1, 2, and 3, respectively. Seroconversion with fIPV im was noninferior to fIPV id for all serotypes. Conclusions. We demonstrated noninferiority of fIPV im compared with id when administered at 4 and 8 months of age. Further investigations in an earlier infant schedule should be pursued to explore fIPV im as option for dose-sparing strategy in countries reluctant to use fIPV id due to programmatic difficulties of id administration.