CAJ | 학술논문

目的利用多种抗戊型肝炎病毒(HEV)IgM、IgG检测试剂对戊型肝炎恢复期血清进行确认,建立戊型肝炎病毒ISG抗体的定量检测方法,初步应用于戊型肝炎疫苗接种后抗体效价检测.方法应用多种抗-HEV IgM、IgG试剂对江苏省确诊的戊型肝炎病人发病后6个月以上的血清样品进行检测;抗-HEV IgG阳性样本应用ORF2 C端抗原和ORF3抗原Western blot进行确认,用WHO抗-HEV Ig标准品标定确认阳性的混合血清抗-HEV IgG的含量,制备抗-HEV IgG定量检测的线性标准品,建立戊型肝炎疫茼免疫后抗体定量检测方法.结果 42份戊型肝炎恢复期血清采用G、K、MP、万泰4种抗-HEV IgG试剂检测的阳性率分别为71.4%、78.6%、92.9%、100%,G试剂阳性率低于MP(X2=5.19,P<0.05)和万泰试剂(χ~2=11.76,P<0.01),K试剂阳性率明显低于万泰试剂(χ~2=7.96,P<0.01);MP、G、X、万泰、K试剂抗-HEV IgM试剂的阳性率分别为21.4%、7.1%、21.4%、64.3%、78.6%;万泰和K试剂的阳性率均明显高于MP(χ~2=15.75,P<0.01;X2=27.43,P<0.01).Western blot确认试验分别有30和18份血清与ORF2、ORF3抗原有阳性反应.13份血清混合为HEV-D01,其抗-HEV IgG浓度经标定为57.94 U/ml.制备了浓度范围为0.077～0.877 U/ml的7个1.5倍系列稀释的定最线性标准品,用于戊肝疫苗临床试验,试验过程中对高、中、低浓度质控品的定量值均在均数±2s范围内,变异系数(cv)分别为16%、16%、12%.结论不同抗-HEV IgM和IgG试剂之间的质量存在较大差别,建立了抗-HEV IsG定量检测标准品,适用于戊肝疫苗的临床血清抗体IgG定量检测.
목적 이용다충항무형간염병독(HEV)IgM、IgG검측시제대무형간염회복기혈청진행학인,건립무형간염병독ISG항체적정량검측방법,초보응용우무형간염역묘접충후항체효개검측.방법 응용다충항-HEV IgM、IgG시제대강소성학진적무형간염병인발병후6개월이상적혈청양품진행검측;항-HEV IgG양성양본응용ORF2 C단항원화ORF3항원Western blot진행학인,용WHO항-HEV Ig표준품표정학인양성적혼합혈청항-HEV IgG적함량,제비항-HEV IgG정량검측적선성표준품,건립무형간염역동면역후항체정량검측방법.결과 42빈무형간염회복기혈청채용G、K、MP、만태4충항-HEV IgG시제검측적양성솔분별위71.4%、78.6%、92.9%、100%,G시제양성솔저우MP(X2=5.19,P<0.05)화만태시제(χ~2=11.76,P<0.01),K시제양성솔명현저우만태시제(χ~2=7.96,P<0.01);MP、G、X、만태、K시제항-HEV IgM시제적양성솔분별위21.4%、7.1%、21.4%、64.3%、78.6%;만태화K시제적양성솔균명현고우MP(χ~2=15.75,P<0.01;X2=27.43,P<0.01).Western blot학인시험분별유30화18빈혈청여ORF2、ORF3항원유양성반응.13빈혈청혼합위HEV-D01,기항-HEV IgG농도경표정위57.94 U/ml.제비료농도범위위0.077～0.877 U/ml적7개1.5배계렬희석적정최선성표준품,용우무간역묘림상시험,시험과정중대고、중、저농도질공품적정량치균재균수±2s범위내,변이계수(cv)분별위16%、16%、12%.결론 불동항-HEV IgM화IgG시제지간적질량존재교대차별,건립료항-HEV IsG정량검측표준품,괄용우무간역묘적림상혈청항체IgG정량검측.
Objective To evaluate anti-HEV IgG and IgM diagnostic kits with sera from convalescent hepatitis E patients and to establish the quantification method of detecting anti-HEV lgG.Methods Detect 42 convalescent serum samples of over 6 months after onset of hepatitis E patients from Jiangsu province with anti-HEV IgM and IgG diagnostic kits. Select and mix the anti-HEV IgG positive sera which were confirmed by Western blot with ORF2 and ORF3 antigen. The mixed serum was calibrated with a WHO anti-HEV Ig standard. A series quantitative linear standard was made for quantitative detection of anti-HEV IgG in hepatitis E vaccine clinical trials phase Ⅲ. Results The positive rates of the anti-HEV IgG di-agnose kits of G, K, MP, Wantai were 71.4%, 78.6%, 92.9% and 100% respectively. The positive rates of G was lower than that of MP (χ~2 = 5.19, P<0.05) and obviously lower than Wantai (χ~2 = 11.76,P<0.01). The positive rates of K was also obviously lower than that of Wantai (χ~2 =7.96, P <0.01).The positive rates of the anti-HEV IgM diagnose kits of MP, G, X, Wantai, K were 21.4%, 7.1%,21.4%, 64.3%, 78.6% respectively. The positive rate of both K and Wantai were obviously higher than that of MP(χ~2 = 15.75 ,P<0.01 ; X2 = 27.43 ,P< 0.01). With the Western blot confirmation test, 30 and 18 sera were reactive to ORF2 and ORF3 antigen separately. The anti-HEV IgG concentration of HEV-D01 mixed by 13 samples was 57.94 U/ml by the calibration. Prepare seven 1.5-fold dilution series of quantita-tive linear standard for HEV vaccine clinical trials phase Ⅲ, concentration range from 0.077 to 0.877 U/ml. The quantitive values of high, medium and low concentrations quality control samples lay in the range of average ± 2s, and the CV of quantitative values were 16%, 16%, 12% respectively. Conclusion The quality of different anti-HEY IgM and IgG diagnose kits were different. This study had set up a set of anti-HEV IgG linear quantitative standard, which fit for detecting anti-HEV IgG antibodies quantitatively in HEVvaccine clinical trial phase Ⅲ.