国际生物制品学杂志
國際生物製品學雜誌
국제생물제품학잡지
INTERNATIONAL JOURNAL OF BIOLOGICALS
2015年
3期
121-126
,共6页
陈蕾%戴新宪%郝春生%张越%宋冬梅%田龙%李秀玲
陳蕾%戴新憲%郝春生%張越%宋鼕梅%田龍%李秀玲
진뢰%대신헌%학춘생%장월%송동매%전룡%리수령
布尼亚病毒科%抗原%酶联免疫吸附测定%方法
佈尼亞病毒科%抗原%酶聯免疫吸附測定%方法
포니아병독과%항원%매련면역흡부측정%방법
Bunyaviridae%Antigens%Enzyme-linked immunosorbent assay%Methods
目的 建立定量检测发热伴血小板减少综合征布尼亚病毒(severe fever with thrombocytopenia syndrome bunyavirus,SFTSV)抗原的方法,以用于疫苗生产过程中SFTSV抗原含量的检测.方法 用SFTSV抗原免疫BALB/c小鼠,制备抗SFTSV单克隆抗体.抗体纯化后以辣根过氧化物酶进行标记,建立双抗体夹心ELISA法.确定该法的线性范围和灵敏度,并对该法的准确性、精密性、专属性进行验证.结果 建立的ELISA法的线性范围为0.117～2.000 mg/L,定量限为0.117 mg/L,线性的决定系数为0.990 3.该法具有良好的准确性和精密度,样品回收率为98％,变异系数均＜8％.该法的专属性良好,对6株不同SFTSV的检测结果均为阳性,而与其他布尼亚病毒、Vero细胞培养上清及其他生产辅料均无交叉反应.结论 成功建立了SFTSV抗原的定量检测方法,为疫苗生产的质量控制奠定了基础.
目的 建立定量檢測髮熱伴血小闆減少綜閤徵佈尼亞病毒(severe fever with thrombocytopenia syndrome bunyavirus,SFTSV)抗原的方法,以用于疫苗生產過程中SFTSV抗原含量的檢測.方法 用SFTSV抗原免疫BALB/c小鼠,製備抗SFTSV單剋隆抗體.抗體純化後以辣根過氧化物酶進行標記,建立雙抗體夾心ELISA法.確定該法的線性範圍和靈敏度,併對該法的準確性、精密性、專屬性進行驗證.結果 建立的ELISA法的線性範圍為0.117～2.000 mg/L,定量限為0.117 mg/L,線性的決定繫數為0.990 3.該法具有良好的準確性和精密度,樣品迴收率為98％,變異繫數均＜8％.該法的專屬性良好,對6株不同SFTSV的檢測結果均為暘性,而與其他佈尼亞病毒、Vero細胞培養上清及其他生產輔料均無交扠反應.結論 成功建立瞭SFTSV抗原的定量檢測方法,為疫苗生產的質量控製奠定瞭基礎.
목적 건립정량검측발열반혈소판감소종합정포니아병독(severe fever with thrombocytopenia syndrome bunyavirus,SFTSV)항원적방법,이용우역묘생산과정중SFTSV항원함량적검측.방법 용SFTSV항원면역BALB/c소서,제비항SFTSV단극륭항체.항체순화후이랄근과양화물매진행표기,건립쌍항체협심ELISA법.학정해법적선성범위화령민도,병대해법적준학성、정밀성、전속성진행험증.결과 건립적ELISA법적선성범위위0.117～2.000 mg/L,정량한위0.117 mg/L,선성적결정계수위0.990 3.해법구유량호적준학성화정밀도,양품회수솔위98％,변이계수균＜8％.해법적전속성량호,대6주불동SFTSV적검측결과균위양성,이여기타포니아병독、Vero세포배양상청급기타생산보료균무교차반응.결론 성공건립료SFTSV항원적정량검측방법,위역묘생산적질량공제전정료기출.
Objective To establish a quantitative detection method for severe fever with thrombocytopenia syndrome bunyavirus (SFTSV) antigen to detect contents of SFTSV antigen during SFTSV vaccine production.Methods Monoclonal antibody was prepared by immunizing BALB/c mouse with SFTSV antigen,and then purified and labeled with horseradish peroxidase to establish a double antibody sandwich ELISA method.Linear range and sensitivity of the ELISA method were detected,and accuracy,precision and specificity of the ELISA method were validated.Results Linear range,quantification limit and determination coefficient of the ELISA method were 0.117-2.000 mg/L,0.117 mg/L and 0.990 3,respectively.Accuracy and precision of the ELISA method were good.Recoveries of internal reference were 98％ and variation coefficient were all lower than 8％.The ELISA method had good specificity.The results for detecting 6 strains of SFTSV with the ELISA method were all positive,and no reactions were observed with other bunyaviruses,Vero cells and other excipients.Conclusion A quantitative detection method for SFTSV antigen is successfully established and lays the foundation for quality control during SFTSV vaccine production.
