狂犬病病毒中和抗体检测试剂盒的效果评价

目的应用快速荧光灶抑制试验(RFFIT)对狂犬病病毒中和抗体检测试剂盒的检测效果进行评价。方法用RFFIT和狂犬病病毒中和抗体检测试剂盒分别对135份人血清和164份动物血清进行狂犬病病毒中和抗体(RVNA)效价检测,应用SPSS22.0分析两种检测方法所得结果的相关性和一致性。结果两种方法所得结果的阳性检出符合率为99.33%(297/299);两种方法对动物血清和人血清的检测结果之间有较好的直线相关关系,相关系数分别为0.897和0.941;同一份标本的重复性检验显示,试剂盒检测方法具有较好的稳定性。结论狂犬病病毒中和抗体检测试剂盒可以用于人和动物血清狂犬病病毒中和抗体的检测。     

新型抗狂犬病病毒核蛋白抗体进行RFFIT检测效果评估

目的评价自主研发的荧光标记抗狂犬病病毒核蛋白单克隆抗体用于快速荧光灶抑制试验(RFFIT)的效果。方法将狂犬病病毒核蛋白的一段线性表位肽与KLH耦联后免疫Balb/c小鼠制备的高效价单克隆抗体抗-RVNP-Mcl,经浓缩、纯化、异硫氰酸荧光素(FITC)标记用于RFFIT检测,以Millipore公司DFA5100为对照试剂,对200例人血清进行平行RFFIT检测,将结果进行统计学分析。结果两种抗体检测结果的线性相关分析显示相关系数(r)=0.980;两种抗体检测定性结果一致率为96.83%,卡方检验显示两种抗体检测定性结果差异无统计学意义(χ2=0.098,P>0.05),发生定性结果不一致的6例(3.00%)血清两种抗体检测结果均小于1 IU/mL;68例两种抗体检测结果均小于或等于1 IU/mL的血清样品中有6例(8.82%)发生结果定性不一致,其余样品均未发生不一致的情况。结论使用荧光标记抗-RVNP-Mcl与DFA5100进行RFFIT检测时结果高度一致,荧光标记抗-RVNP-Mc1可供RFFIT检测使用。     

改良抗体结合试验的建立和在灭活狂犬病疫苗效价检测中的应用

目的对抗体结合试验(ABT)进行改进并将其用于灭活狂犬病疫苗效力的测定。方法将快速荧光灶抑制试验(RFFIT)与ABT方案融合,在疫苗样品稀释、剩余中和抗体检测、结果计算方面进行改进,形成改良抗体结合试验(M-ABT);应用M-ABT对10种灭活狂犬病疫苗随库存时间增加疫苗效力改变的情况进行检测。结果 M-ABT可以在28 h内完成,比ABT节省2 d,检测结果与人用狂犬病疫苗效价测定法(NIH)基本等效;10种疫苗随着库存时间增加疫苗效力下降。结论 M-ABT法能够成功建立,并用于灭活狂犬病疫苗效力检测效果较满意。     

狂犬病疫苗接种者抗狂犬病病毒中和抗体影响因素分析

目的探讨狂犬病疫苗接种后体内抗狂犬病病毒中和抗体产生的影响因素。方法采集2018年1-8月山东、江苏、河南、河北、湖南和安徽6省疑似狂犬病Ⅱ/Ⅲ级暴露后接种狂犬病疫苗的暴露者血清标本,快速荧光灶抑制实验(RFFIT)检测抗狂犬病病毒中和抗体(RVNA)水平,采用SPSS 19.0软件进行数据统计分析。结果全程接种狂犬病疫苗的血清样本为498份,RFFIT检测结果显示,RVNA几何平均滴度(GMT)为4.94 IU/ml,血清阳性率为96.18%(479/498)。不同组别抗体阳性率分析结果显示,免疫时间各组阳性率之间差异有统计学意义(χ^2=7.888,P=0.039),30~90 d组抗体阳性率高于91~180 d组,其他各组比较差异均无统计学意义。多因素分析结果显示年龄、免疫时间是RVNA水平的重要影响因素,其中年龄与RVNA水平呈正相关,免疫时间与RVNA水平呈负相关。免疫时间对RVNA水平的影响最大,影响重要性为0.62。结论接种狂犬病疫苗后能产生可靠的免疫效果,年龄、免疫时间等因素对抗体阳性率及RVNA水平有一定影响。暴露后在及时进行规范完整的暴露后处置外,应对经常暴露于狂犬病的高危人群、免疫力低下者或患有免疫抑制性疾病的人群定期检测血清抗体,对于RVNA滴度小于0.5 IU/ml的接种者应及时进行加强免疫,补接种疫苗,从而有效预防狂犬病。     

狂犬病毒核蛋白单抗和荧光抗体中和试验的建立与应用

目的建立狂犬病毒血清中和抗体效价的快速检测方法，以实现狂犬疫苗免疫效果的定量评价，指导高危人群及动物种群的免疫预防。方法采用常规方法建立杂交瘤细胞株并制备抗狂犬病毒核蛋白单抗，以亲和层析法进行纯化，异硫氰酸荧光黄标记制备荧光抗体；以BHK-21细胞系培养狂犬病毒细胞适应株CVS-11，以染毒细胞测定荧光抗体的工作浓度后，通过荧光抗体染色法测定CVS-11细胞半数感染量（TCID50）；利用狂犬病毒灭活疫苗免疫犬制备抗狂犬病毒血清，并以国际兽疫局标准品进行标定；参考世界卫生组织及国际兽疫局推荐方法，确定中和试验程序，对51份样品血清进行测定，并与国际狂犬病参考实验室的检测结果进行比较，评价其精确及可靠程度。以Kaber法公式和Microsoft Excel软件为基础，设计被测样品中和效价计算方法。结果制备的抗狂犬病毒核蛋白单抗的亚型为IgG2a；纯化、标记后的荧光抗体工作浓度为1：400；CVS-11株的使用量为每孔100 TCID50；本研究建立的检测程序对51份人及犬血清测得的中和效价结果与国际参考实验室的测定结果一致；自行设计的计算方法操作简便，计算快捷。结论成功建立了与国际标准相当的狂犬病毒中和抗体效价测定和计算方法。     

微量细胞中和试验和空斑减少中和试验在HFRSV中和抗体测定中的比较

为了满足肾综合征出血热(HFRS)血清流行病学、病毒分型和疫苗研究的需要,国内外已相继建立了多种 HFRS 病毒(HFRSV)中和抗体的测定方法,其中应用较多的是空斑减少中和试验(PRNT)、微量细胞中和试验。从测定的灵敏度和准确性考虑,PRNT无疑具有更多的优点,但是鉴于目前国内的实验条件,HFRSV 的 PRNT 尚不能普遍开展,微量细胞中和试     

人巨细胞病毒中和抗体快速微量检测方法的建立及应用

目的建立具有较好试验重复性的人巨细胞病毒中和抗体快速微量检测方法。方法建立快速微量中和试验方法,用于健康人血浆及IVIG中的CMV中和抗体效价检测。结果用该方法检测336份健康人血浆,其中CMV中和抗体效价≥1∶128的比例为28.1%;检测27批"蓉生静丙"和9批国内其它厂家IVIG的CMV中和抗体效价,其CMV中和抗体效价的GMTs分别为1∶498和1∶485。结论建立了快速、简便、检测通量高、重复性较好的快速微量中和试验方法,为CMV-IVIG的研制提供了可供选择的血浆筛选方法。     

山西和河南省部分地区人群流行性乙型脑炎病毒中和抗体水平调查

目的调查山西和河南省部分地区健康人群血清中流行性乙型脑炎(乙脑)病毒中和抗体水平。方法对采集的血清标本按1:10,1:20,1:40,1:80四个稀释度稀释后进行乙脑病毒中和试验,结果判定采用细胞病变观察方法并结合ELISA测定结果进行综合判定。结果调查结果显示,所测标本中乙脑病毒中和抗体保护率以临猗县最高(87.6%),其次是栾川县(58.2%)和万荣县(51.4%)。经χ2检验,临猗县与万荣县中和抗体保护率差异有统计学意义(χ2=42.812,P=0.000),临猗县的中和抗体保护率高于万荣县,栾川县与万荣县的中和抗体保护率差异无统计学意义(χ2=1.466,P=0.226)。两个年龄组(≤40岁和40岁)中和抗体保护率比较显示,临猗县差异无统计学意义(χ2=0.057,P=0.811),万荣县差异有统计学意义(χ2=5.505,P=0.019),栾川县差异无统计学意义(χ2=2.129,P=0.145)。结论三个县健康人群乙脑病毒中和抗体保护率不同,这可能与当地乙脑的流行状况以及疫苗的接种情况有关。     

两种联合免疫方式下血清狂犬病病毒抗体结果比较

目的比较头面部和手足末端暴露者在两种不同的联合免疫方式下,血清狂犬病病毒抗体的差异性。方法选择2010年4月至2013年6月期间来江西省赣州市疾病预防控制中心就诊的头面部、手足末端Ⅲ级暴露并同意使用狂犬疫苗和狂犬病人免疫球蛋白fHumanrabiesimmunoglobulin。HRIG）联合免疫的患者,随机分为实验组和对照组,实验组采用HRIG分三步给药的新注射法．随后按0、7、21程序2—1—1四针法接种狂犬疫苗;对照组采用常规法注射HRIG,随后按O、3、7、14、28程序五针法接种疫苗。分别在两组患者首针疫苗后第7d（D7）~ll第45d（D。）采血清,酶联免疫法测狂犬病病毒IgG抗体,比较两组抗体阳性率差异。结果实验组D,的狂犬病病毒IgG抗体阳性率明显高于对照组（P〈0．01）,而D。两组无明显差异p0．05）。结论新联合免疫方式能提高免疫早期的抗体水平并保持后期抗体不受影响。     

汕头地区人群中埃可病毒抗体的检测

目的 :观察人群中感染埃可病毒 (ECHO)状况。方法 :采用中和抗体试验测定人群血清抗体 ,血清作1∶10～ 1∶3 2 0稀释 ,病毒用 10 0TCID50 在微量培养板上进行 ;ECHO病毒 6个型分别与血清等量混合 ,结合 1h ,接种细胞 (同时设立病毒、细胞对照 ) ,观察细胞病变。结果 :血清ECHO 7、9、14、15、2 4、2 7型中和抗体阳性率和几何平均滴度依次为 :2 4 2 %和 7 18,16 0 %和 6 3 7,3 8 9%和 9 42 ,19 9%和 6 3 7,15 7%和 6 3 4,10 8%和 5 64。其中<3岁年龄组的抗体阳性率较低 ,为 2 6%～ 3 2 5 % ,15～ 40岁年龄组抗体阳性率最高 ,为 11 3 %～ 5 2 8%。结论 :汕头地区人群中ECHO 7、9、14、15、2 4、2 7型病毒的传播广泛 ,以青壮年感染率较高 ,婴幼儿较低 ,可能与参加社会活动多少有关。     

定量检测人狂犬病毒中和抗体间接酶联免疫吸附测定方法的建立

目的 建立定量检测人狂犬病毒中和抗体间接酶联免疫吸附测定(ELISA)方法.方法 选择包被抗原和酶标二抗的最适浓度,用人狂犬病毒抗体标准品标化系列工作标准品,对临床收集的各类血清标本进行检测和分析.结果 间接ELISA定量检测中和抗体的最佳包被糖蛋白抗原浓度为10μg/L,酶标二抗的工作浓度为1:4 000,临床检验,间接ELISA定量方法灵敏度为100Vo.特异度为98.2%,总符合率99.0%;与快速荧光灶抑制实验呈(RFFIT)高度相关性(r=0.981).结论 本研究建立的定量检测狂犬病中和抗体方法,可以用于人狂犬病毒中和抗体的定量检测.     

Binding and neutralizing anti-AAV antibodies: Detection and implications for rAAV-mediated gene therapy

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砷快速检测盒与原子荧光法测定水砷含量结果对比

目的　比较水砷快速检测试剂盒(速测法)和氢化物发生-原子荧光光谱法(HG-AFS)测定饮水中砷含量的可靠性和应用价值。方法　用速测法和HG-AFS测定水样747份,将2种方法检测结果进行对比,数据处理应用SPSS 10.0软件进行配对设计的秩和检验。结果　2种方法测得的结果总体差异无统计学意义(u=1.32,P0.05),但速测法结果在0.03～0.05 mg/L 之间与HG-AFS结果差异有统计学意义(u=2.42,P0.05)。结论　HG-AFS稳定可靠,速测法操作简单,可以在现场应用。用速测法初步筛选,用HG-AFS对速测法结果含砷定量分析,既可减少工作量,又能保证分析结果的可靠性,完全能满足地方性砷中毒工作的需求。     

Performance evaluation of four rapid antibody tests for the detection of severe acute respiratory syndrome coronavirus 2

BackgroundThe prompt detection of severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) is important in the therapeutic management of infected patients. Rapid diagnostic tests are widely used for this purpose. This study aimed to evaluate the clinical performance of four SARS‐CoV‐2 immunoglobulin IgG/IgM rapid diagnostic tests in the detection of SARS‐CoV‐2.MethodsNasopharyngeal and oropharyngeal swabs and/or sputum were collected from 30 patients infected with SARS‐CoV‐2 and 30 healthy volunteers. All specimens were tested using four SARS‐CoV‐2 IgG/IgM rapid diagnostic tests and real‐time polymerase chain reaction. We assessed the clinical sensitivity and specificity of the tests.ResultsThe clinical sensitivity of FREND™, SsmarTest™, BIOCREDIT™, and IVDLAB™ was 96.67%, 100.00%, 100.00%, and 96.67%, respectively, compared to real‐time polymerase chain reaction. The clinical specificity was 96.67%, 100.00%, 86.67%, and 96.67%, respectively.ConclusionThese findings could expedite the detection of SARS‐CoV‐2 and thus reduce the risk of further transmission of the virus.     

Performance evaluation of three automated quantitative immunoassays and their correlation with a surrogate virus neutralization test in coronavirus disease 19 patients and pre‐pandemic controls

BackgroundSARS‐CoV‐2 pandemic is currently ongoing, meanwhile vaccinations are rapidly underway in some countries. The quantitative immunoassays detecting antibodies against spike antigen of SARS‐CoV‐2 have been developed based on the findings that they have a better correlation with the neutralizing antibody.MethodsThe performances of the Abbott Architect SARS‐CoV‐2 IgG II Quant, DiaSorin LIAISON SARS‐CoV‐2 TrimericS IgG, and Roche Elecsys anti‐SARS‐CoV‐2 S were evaluated on 173 sera from 126 SARS‐CoV‐2 patients and 151 pre‐pandemic sera. Their correlations with GenScript cPass SARS‐CoV‐2 Neutralization Antibody Detection Kit were also analyzed on 173 sera from 126 SARS‐CoV‐2 patients.ResultsArchitect SARS‐CoV‐2 IgG II Quant and Elecsys anti‐SARS‐CoV‐2 S showed the highest overall sensitivity (96.0%), followed by LIAISON SARS‐CoV‐2 TrimericS IgG (93.6%). The specificities of Elecsys anti‐SARS‐CoV‐2 S and LIAISON SARS‐CoV‐2 TrimericS IgG were 100.0%, followed by Architect SARS‐CoV‐2 IgG II Quant (99.3%). Regarding the correlation with cPass neutralization antibody assay, LIAISON SARS‐CoV‐2 TrimericS IgG showed the best correlation (Spearman rho = 0.88), followed by Architect SARS‐CoV‐2 IgG II Quant and Elecsys anti‐SARS‐CoV‐2 S (all rho = 0.87).ConclusionsThe three automated quantitative immunoassays showed good diagnostic performance and strong correlations with neutralization antibodies. These assays will be useful in diagnostic assistance, evaluating the response to vaccination, and the assessment of herd immunity in the future.     

Comparison of four enzyme immunoassays for the detection of immunoglobulin G antibody against glomerular basement membrane

Goodpasture syndrome is a life-threatening autoimmune kidney and pulmonary disease that is characterized by pulmonary hemorrhage, renal failure, and the presence of autoantibodies against glomerular basement membrane (GBM) by indirect fluorescent antibody (IFA) techniques. In 1988, these antibodies were found to be specific for the noncollagen region of the alpha3 collagen IV chain. The antigen is characterized by a restricted tissue distribution occurring mainly in the kidneys and lungs. Specific enzyme immunoassays (EIAs) for anti-GBM have now been developed for in vitro diagnostic use in the laboratory. Our objective in this study was to compare the results obtained using four different EIAs for detecting anti-GBM IgG antibody with those using the standard IFA method using tissue from human kidney. Thirty-two patients with suspected Goodpasture syndrome, and 10 control sera were included in the study. GBM EIAs were purchased from or donated by the following vendors: Scimedx Corporation (Denville, NJ), INOVA Diagnostics (San Diego, CA), The Binding Site (Birmingham, England), and Wieslab (distributed by DiaSorin, Inc., Stillwater, MN). Percent agreement, sensitivity, and specificity were calculated for each EIA as compared to IFA. The results were as follows: Binding Site: 83.3, 100.0, and 72.0%; Wieslab: 95.2, 94.1, and 96.0%; INOVA: 96.2, 100.0, and 92.3%; and Scimedx: 81.0, 100.0, and 68.0%. We conclude that the INOVA and Wieslab GBM IgG EIAs compared well with the standard GBM IFA method using tissue from human kidney. The Scimedx and Binding Site EIAs had eight and seven false-positive results, respectively, when compared to GBM IFA using human kidney. The authors recommend conducting thorough evaluations of EIAs that screen for anti-GBM antibody before their implementation in the clinical laboratory. We also recommend confirming GBM EIA-positive sera by the standard IFA method using tissue from human kidney.     

化学发光酶免疫分析法及ELISA检测抗-HCV的结果比较

目的对比化学发光酶免疫分析法及酶联免疫吸附试验(ELISA)用于丙型肝炎抗体检测的效果。方法选取丙型肝炎患者140例,抽取血液标本后分别进行丙型肝炎病毒(HCV)相关抗体的化学发光酶免疫分析法及ELISA法检测。结果化学发光酶免疫分析法检出抗-HCV阳性的检出率为98.6%,高于ELISA法的检出率(94.3%),差异有统计学意义(P0.05)。化学发光酶免疫分析法对于抗AMA-M2抗体、抗3E抗体、抗SP100抗体、抗PML抗体、抗GP210抗体的检测阳性率分别为80.0%、73.6%、37.9%、55.7%和47.9%,而ELISA法检测结果分别为12.9%、12.9%、13.6%、10.7%和6.4%,差异均有统计学意义(P0.05)。结论相对于ELISA法,化学发光酶免疫分析法在丙型肝炎中的应用有较高的检出阳性率,对于相关抗体检测有较高的敏感性,值得推广应用。     

Comparison of three microtube column agglutination systems for antibody screening: DG Gel, DiaMed-ID and Ortho BioVue

The aims of the present study were to evaluate the estimated diagnostic accuracy of a new microtube column agglutination system (DG Gel, Diagnostic Grifols, Barcelona, Spain), to analyse the antibody reactivity and to compare the data with the two well-established DiaMed-ID and Ortho BioVue systems. We collected 3024 consecutive samples from blood donors, transfusion recipients and pregnant women, and 100 samples containing antibodies of known specificity. All these samples were tested in parallel by the three microtube agglutination systems. The estimated sensitivity was 100% for DG Gel and Ortho BioVue and 97.58% for DiaMed-ID. The estimated specificity was 99.93% for Ortho BioVue and 100% for DiaMed-ID and DG Gel. The score mean and range of the antibody titration of DG Gel, DiaMed-ID and Ortho BioVue were 34.31 (5-119), 30.3 (3-121) and 37.38 (3-112), respectively. All three column agglutination systems work well showing a high estimated diagnostic accuracy.