用单克隆抗体分析鉴定登革3型病毒的不同抗原决定簇

登革病毒(DEN)抗原成分复杂,与许多虫媒披膜病毒都有严重的血清学交叉反应。杂交瘤单克隆抗体(Mcab)技术建立之后,Dittmar等人首先制备出了抗DEN-3病毒的McAb,可进行DEN-3病毒的鉴定分型。随后,Henchal等人制备出了抗DEN1-4型病毒的McAb,用这些McAb进行DEN病毒抗原决定簇研究,依据间接免疫荧光染色(IFA)证实了DEN病毒的四种不同抗原决定簇,即:黄病毒属特异的、DEN-1-4型病毒特异的、DEN-1、3型病毒特异的和DEN病毒各型型特异的。阎国珍等人在研究抗DEN-4病毒McAb时,获得了抗DEN-2、4型病毒特异的McAb,证明DEN-2和DEN-4病毒间有共同特异抗原决定簇。本文介绍通过用抗DEN-3病毒McAb对不同披膜病毒抗原交叉反应测定来分析鉴定     

我国几种蚊虫对登革病毒媒介效能的实验研究

本文以蚊体感染率和涎腺感染率为指标,比较了我国6株白纹伊蚊(Aedesalbopictus)和海南株埃及伊蚊(Ae aegypti)对登革2(DEN-2)型病毒的媒介效能。埃及伊蚊的涎腺感染率显著高于各株白纹伊蚊的总涎腺感染率。蚊体感染率则以上海株白纹伊蚊最高,福州株次之。用群体叮吸细胞液的方法首次证明了经口感染的黄斑伊蚊(Ae Jlavopictus)经过15天外潜伏期后,可传播登革病毒。致倦厙蚊(Culex pipiens quinquefasciatus)(广州株)吸入含有DEN-2型病毒的混合液后,随着时间的延长,阳性率迅速下降,感染后3天和21天分离,均为阴性。     

应用酶标抗体染色快速检查微量细胞培养中的病毒抗原

我们应用微量细胞培养作病毒诊检,在病毒性肺炎及登革热的病原诊断以及在鼻病毒的定型等工作中都取得较满意的结果。但因采用细胞病变作为判别结果指标不能快速作出诊断,为此,我们采用酶标抗体染色法试行检出微量细胞培养中的病毒抗原,现将所得结果予以报道。材料与方法 1.病毒森林脑炎病毒(TBE)森张株,乳鼠脑(SM B)11代;登革病毒2型(DEN-2)新几内亚B株,SMB11代;乙型脑炎病毒(JBE)高株,SMB12代,均为本室保存的毒种,本试验应用10％鼠脑悬液或BHK-13细胞中传1～2代的病毒液。甲型流     

肾综合征出血热病毒单克隆抗体杂交瘤细胞系的建立与鉴定

用陕西省分离的肾综合征出血热(HFRS)病毒82-010H株免疫BALB/c纯系鼠和CxS/2小鼠,取免疫鼠脾细胞与SP2/O小鼠骨髓瘤细胞融合,获得10株分泌HFRS病毒特异性抗体的杂交瘤细胞系,对其中3H_4和4B_9两株单克隆抗体进行了鉴定。用各地分离的16株HFRS病毒抗原片与该两株单克隆抗体作了间接免疫荧光试验,结果表明:4B_9单克隆抗体只能与重型(黑线姬鼠型)HFRS疫区的病毒呈现阳性免疫荧光反应,而对轻型(褐家鼠及大林姬鼠型)HFRS疫区分离的病毒株不呈现免疫荧光反应;而3H_4单克隆抗体又能将重型和轻型HFRS毒株再行区分,这可能对HFRS病毒血清学分型和抗原分析以及流行病学调查研究等均有意义。     

登革病毒单克隆抗体鉴定及其血清学性质观察

利用补体结合试验(CF)和血凝抑制试验(HI)对83份免疫荧光阳性的登革病毒单克隆抗体进行了鉴定,11份CF阳性,8份HI阳性。登革1型和4型病毒的单克隆抗体仅有CF活性,其中4F_7和4F_9滴度达到1:20480～1:40960,并且为CF型特异。而登革2型和3型病毒的单克隆抗体,则仅有HI活性,除3D_4为HI型特异外,其余均为HI属特异,与其他黄病毒属抗原有明显交叉。     

登革2型病毒单克隆抗体的制备与鉴定

用登革2型病毒参考株免疫的BALB/C小鼠脾细胞与SP2/0小鼠骨髓瘤细胞在PEG1000作用下进行融合,获得了10株分泌抗登革2型病毒单克隆抗体的杂交瘤。经间接免疫荧光鉴定,有4株杂交瘤产生的单克隆抗体对登革2型病毒是型特异的;2株对登革1型有交叉反应;1株对登革3型有交叉反应;1株对登革4型有交叉反应;另2株为黄病毒属特异的。经补体结合和血凝抑制试验鉴定,有2个单克隆抗体是黄病毒属血凝抑制特异的。这10个单克隆抗体对登革热的诊断是很有用的。     

利用DNA免疫建立分泌登革2型病毒E蛋白单抗的杂交瘤细胞系

目的:利用DNA免疫建立分泌登革2型病毒E蛋白特异单抗的杂交瘤细胞系,为研究E蛋白的结构和功能及其抗原表位提供新手段.方法:以构建的病毒全长prM-E基因的真核重组质粒DNA作为免疫原,免疫BALB/c小鼠后将其脾细胞和SP2/0瘤细胞融合,通过IFA、间接ELISA和空斑减少中和试验对杂交瘤细胞系进行筛选和鉴定.结果:获得了4株分泌登革2型病毒E蛋白单抗的杂交瘤细胞系(2B4,6B4,4C10和2D5),它们结合的抗原表位均位于E蛋白Ⅲ区,其中4C10对登革2型病毒具有中和活性.这些单抗与登革1、3和4型病毒有强的交叉反应,但与黄病毒其他成员反应较弱.结论:DNA免疫法可用于分泌登革2型病毒E蛋白特异单抗的杂交瘤细胞系的建立,该结果有利于登革病毒E蛋白特异抗原表位的研究及新的登革病毒诊断试剂的研制.     

登革1型病毒的单克隆抗体

我们用SP2/0-Ag14小鼠骨髓瘤细胞与用登革1型病毒免疫的BALB/C小鼠脾细胞在融合剂PEG-1000的诱导下进行融合,获得了15株产生抗登革1型病毒单克隆抗体的杂交瘤细胞系。其中14株属登革病毒型特异,1株属登革病毒亚群特异,它们的荧光效价均在10,240以上。15株杂交瘤均无血凝活性,3株有补体结合活性。1D_4,1B_9的IgG亚类为IgG_1,1F_(11)为IgG_2a,轻链均为K型。1株杂交瘤(1D_4)的染色体数为87～115。用SDS-PAGE测定1D_4分子量,重链为54,000,轻链为24,000。15株杂交瘤连续传代3～6个月,冻存复苏后抗体水平未见下降。     

森林脑炎病毒单克隆抗体的制备与鉴定

用森林脑炎病毒森张株免疫的BALB/c小鼠脾细胞与小鼠骨髓瘤细胞在融合剂PEG-1000的作用下进行融合,用间接免疫荧光法筛选,获得了6株分泌抗森林脑炎病毒单克隆抗体的杂交瘤细胞株.通过免疫荧光交叉鉴定,6份单克隆抗体均与黄病毒属的登革4型病毒、流行性乙型脑炎病毒无属间交叉.与森林脑炎病毒COφ株、国内的森候株、森董株有交叉反应,属森林脑炎病毒种特异.6份单克隆抗体均无血凝抑制活性和补体结合活性.3株杂交瘤细胞培养上清浓缩物经抗鼠Ig免疫血清双扩鉴定为IgM.染色体数的检查为89～110条.6株杂交瘤细胞连续传代3～6个月,冻存复苏后仍能稳定地分泌特异的单克隆抗体.     

脾内注射法免疫小鼠制备抗—HBc单克隆抗体的研究

用基因工程菌产乙型肝炎核心抗原(HBcAg)对 Balb/c 小鼠脾内多点注射法免疫,将该鼠的脾淋巴细胞与鼠骨髓瘤细胞在 PEG 作用下融合,用固相放射免疫分析法(SPRIA)筛选,成功地获得了2株抗-HBc 阳性的杂交瘤细胞株。通过中和试验和交叉鉴定,2株抗-HBc 单克隆抗体只能与 HBcAg 起反应,而不能与 HBsAg 和 HBeAg 反应。经多次亚克隆后,所得腹水中的抗-HBc 滴度达1:64000～1:28000。这2株单克隆抗体做包被抗体检测 HBcAg 和(125)~Ⅰ标记查患者血清标本时,所测得的结果与人抗-HBc 多克隆抗体做包被或(125)~Ⅰ标记所测得的结果相一致。2株杂交瘤细胞连续传代3个多月,冻存复苏后仍能稳定地分泌特异的抗-HBc 单克隆抗体。     

Knee injury and Osteoarthritis Outcome Score (KOOS) - validation of a Swedish version

The Knee injury and Osteoarthritis Outcome Score (KOOS) is a self-administered instrument measuring outcome after knee injury at impairment, disability, and handicap level in five subscales. Reliability, validity, and responsiveness of a Swedish version was assessed in 142 patients who underwent arthroscopy because of injury to the menisci, anterior cruciate ligament, or cartilage of the knee. The clinimetric properties were found to be good and comparable to the American version of the KOOS. Comparison to the Short Form-36 and the Lysholm knee scoring scale revealed expected correlations and construct validity. Item by item, symptoms and functional limitations were compared between diagnostic groups. High responsiveness was found three months after arthroscopic partial meniscectomy for all subscales but Activities of Daily Living.     

Endovascular management of carotid-cavernous fistulas

Objective To investigate endovascular treatment of traumatic direct carotid-cavernous fistulas （CCF） and their complications such as pseudoaneurysms. Methods： Over a five-year period, 22 patients with traumatic direct CCFs were treated endovascularly in our institution. Thirteen patients were treated once with the result of CCF occluded, 8 twice and 1 three times. Treatment modalities included balloon occlusion of the CCF, sacrifice of the ipsilateral internal carotid artery with detachable balloon, coll embolization of the cavernous sinus and secondary pseudoaneurysms, and covered-stem management of the pseudoaneurysms. Results All the direct CCFs were successfully managed endovascularly. Four patients developed a pseudoaneurysm after the occlusion of the CCF with an incidence of pseudoaneurysm formation of 18.2% （4/22）. A total number of 8 patients experienced permanent occlusion of the ICA with a rate of ICA occlusion reaching 36.4% （8/22）. Followed up through telephone consultation from 6 months to 5 years, all did well with no recurrence of CCF symptoms and signs. Conclusion Traumatic direct CCFs can be successfully managed with endovascular means. The pseudoaneurysms secondary to the occlusion of the CCFs can be occluded with stent-assisted coiling and implantation of covered stents.     

The acutely limping child

Acute limping may be the result of multiple pathologies in children. The differential diagnosis varies based on the age of the child. Irrespective of age, the initial imaging work-up includes AP and frog leg radiographs of the pelvis and ultrasound; MRI may sometimes be helpful. In children less than 3 years, infections and trauma are most frequent. MRI is the imaging modality of choice when osteomyelitis is clinically suspected. Between the ages of 3 and 10 years, transient synovitis of the hip and Legg-Calvé-Perthes disease are main considerations but infection, inflammation and focal bony lesions are also considered. In children over 10 years, slipped capital femoral epiphysis also is considered.     

Do Balance Board Training Programs Reduce the Risk of Ankle Sprains in Athletes?

Introduction Ankle sprains are the most common musculo-skeletal injury that occurs in athletes,particularly in sports that require jumping and landing on one foot such as soccer,and basketball(1-4).These injuries often result in significant time loss from participation,long-term disability,and have a major impact on health care costs and resources(5-8).     

High Intensity Interval Training:New Insights

KEY POINTS ·High-intensity interval training(HIT)is characterized by repeated sessions of relatively brief，intermittent exercise.often performed with an“a11 out”effort or at an intensity close to that which elicits peak oxygen uptake(i.e.，≥90%of VO2 peak).     

Developmental validation of the PowerPlex(®) ESI 16 and PowerPlex(®) ESI 17 Systems: STR multiplexes for the new European standard

In response to the ENFSI and EDNAP groups’ call for new STR multiplexes for Europe, Promega^® developed a suite of four new DNA profiling kits. This paper describes the developmental validation study performed on the PowerPlex^® ESI 16 (European Standard Investigator 16) and the PowerPlex^® ESI 17 Systems. The PowerPlex^® ESI 16 System combines the 11 loci compatible with the UK National DNA Database^®, contained within the AmpFlSTR^® SGM Plus^® PCR Amplification Kit, with five additional loci: D2S441, D10S1248, D22S1045, D1S1656 and D12S391. The multiplex was designed to reduce the amplicon size of the loci found in the AmpFlSTR^® SGM Plus^® kit. This design facilitates increased robustness and amplification success for the loci used in the national DNA databases created in many countries, when analyzing degraded DNA samples. The PowerPlex^® ESI 17 System amplifies the same loci as the PowerPlex^® ESI 16 System, but with the addition of a primer pair for the SE33 locus. Tests were designed to address the developmental validation guidelines issued by the Scientific Working Group on DNA Analysis Methods (SWGDAM), and those of the DNA Advisory Board (DAB). Samples processed include DNA mixtures, PCR reactions spiked with inhibitors, a sensitivity series, and 306 United Kingdom donor samples to determine concordance with data generated with the AmpFlSTR^® SGM Plus^® kit. Allele frequencies from 242 white Caucasian samples collected in the United Kingdom are also presented. The PowerPlex^® ESI 16 and ESI 17 Systems are robust and sensitive tools, suitable for the analysis of forensic DNA samples. Full profiles were routinely observed with 62.5 pg of a fully heterozygous single source DNA template. This high level of sensitivity was found to impact on mixture analyses, where 54–86% of unique minor contributor alleles were routinely observed in a 1:19 mixture ratio. Improved sensitivity combined with the robustness afforded by smaller amplicons has substantially improved the quantity of data obtained from degraded samples, and the improved chemistry confers exceptional tolerance to high levels of laboratory prepared inhibitors.