国际流行病学传染病学杂志
國際流行病學傳染病學雜誌
국제류행병학전염병학잡지
INTERNATIONAL JOURNAL OF EPIDEMIOLOGY AND INFECTIOUS DISEASE
2014年
4期
249-251
,共3页
肝炎病毒,甲型%灭活疫苗%解离液%铝佐剂%ELISA
肝炎病毒,甲型%滅活疫苗%解離液%鋁佐劑%ELISA
간염병독,갑형%멸활역묘%해리액%려좌제%ELISA
Hepatitis A virus%Inactivated vaccine%Dissociation buffer%Aluminium Adjuvant%ELISA
目的 建立铝吸附甲型肝炎(甲肝)灭活疫苗中甲肝病毒(HAV)抗原的解离方法.方法 选择20％二乙醇胺1.25 mL、10％Triton X-100 0.2 mL和PBS 8.55 mL混合液作为解离液A;pH7.2的3％(w/v)柠檬酸缓冲液作为解离液B;20％(w/v)柠檬酸三钠为解离液C.分别检测3种解离液在不同的解离条件下对铝佐剂甲型肝炎灭活疫苗的解离情况,用ELISA法检测解离后的HAV抗原含量,计算抗原回收率,确定最佳解离液及解离条件,并验证其可重复性.结果 3种解离液HAV抗原解离试验显示,解离液C在室温(20～28℃)条件下解离1h,HAV抗原的回收率最高,可达95％及以上,重复试验的变异系数均＜10％.结论 20％ (w/v)柠檬酸三钠可作为铝佐剂甲肝灭活疫苗的首选抗原解离剂,该解离剂配方简单,操作简便,结果重复性好,准确度高.
目的 建立鋁吸附甲型肝炎(甲肝)滅活疫苗中甲肝病毒(HAV)抗原的解離方法.方法 選擇20％二乙醇胺1.25 mL、10％Triton X-100 0.2 mL和PBS 8.55 mL混閤液作為解離液A;pH7.2的3％(w/v)檸檬痠緩遲液作為解離液B;20％(w/v)檸檬痠三鈉為解離液C.分彆檢測3種解離液在不同的解離條件下對鋁佐劑甲型肝炎滅活疫苗的解離情況,用ELISA法檢測解離後的HAV抗原含量,計算抗原迴收率,確定最佳解離液及解離條件,併驗證其可重複性.結果 3種解離液HAV抗原解離試驗顯示,解離液C在室溫(20～28℃)條件下解離1h,HAV抗原的迴收率最高,可達95％及以上,重複試驗的變異繫數均＜10％.結論 20％ (w/v)檸檬痠三鈉可作為鋁佐劑甲肝滅活疫苗的首選抗原解離劑,該解離劑配方簡單,操作簡便,結果重複性好,準確度高.
목적 건립려흡부갑형간염(갑간)멸활역묘중갑간병독(HAV)항원적해리방법.방법 선택20％이을순알1.25 mL、10％Triton X-100 0.2 mL화PBS 8.55 mL혼합액작위해리액A;pH7.2적3％(w/v)저몽산완충액작위해리액B;20％(w/v)저몽산삼납위해리액C.분별검측3충해리액재불동적해리조건하대려좌제갑형간염멸활역묘적해리정황,용ELISA법검측해리후적HAV항원함량,계산항원회수솔,학정최가해리액급해리조건,병험증기가중복성.결과 3충해리액HAV항원해리시험현시,해리액C재실온(20～28℃)조건하해리1h,HAV항원적회수솔최고,가체95％급이상,중복시험적변이계수균＜10％.결론 20％ (w/v)저몽산삼납가작위려좌제갑간멸활역묘적수선항원해리제,해해리제배방간단,조작간편,결과중복성호,준학도고.
Objective To develop a method for antigen dissociation and content determination of hepatitis A virus (HAV) inactivated vaccine containing aluminium adjuvant.Methods Buffer A:20％ diethanolamine 1.25 mL,10％ Triton X-100 0.2 mL,and PBS 8.55 mL; buffer B:3％(w/v) citric acid with pH=7.2; buffer C:20％(w/v) trisodium citrate.The above 3 buffers were used to dissociate HAV inactivated vaccine containing aluminium adjuvant in various conditions.The content of antigen of those resultant samples was determined by ELISA,and then the recovery rate was calculated to determine the best buffer and conditions of dissociation.The repeatability of established method was verified.Results Dissociation test showed that when HAV inactivated vaccine containing aluminium adjuvant was treated with buffer C at 20-28 ℃ for 1 hour,the recovery rate was the highest of 95％ or above.Coefficient of variation of repeated trials was less than 10％.Conclusions 20％(w/v) trisodium citrate is the best buffer for dissociating antigen from HAV inactivated vaccine containing aluminium adjuvant.The buffer is easy to prepare and operate with good reproducibility and high accuracy.
