中华实验和临床病毒学杂志
中華實驗和臨床病毒學雜誌
중화실험화림상병독학잡지
CHINESE JOURNAL OF EXPERIMENTAL AND CLINICAL VIROLOGY
2013年
5期
382-384
,共3页
侯俊%郭桐生%赵静%刘爱霞%宋永继%郭静霞%刘佳%陈霖%徐军
侯俊%郭桐生%趙靜%劉愛霞%宋永繼%郭靜霞%劉佳%陳霖%徐軍
후준%곽동생%조정%류애하%송영계%곽정하%류가%진림%서군
酶联免疫吸附测定%高尔基体%肝肿瘤
酶聯免疫吸附測定%高爾基體%肝腫瘤
매련면역흡부측정%고이기체%간종류
Enzyme-linked immunosorbent assay%Golgi apparetus%Liver neoplasms
目的 建立人血清中高尔基体蛋白73(GP73)的定量酶联免疫吸附试验(ELISA)检测方法.方法 辣根过氧化物酶标记GP73单抗,优化各类反应液的工作浓度和各种反应条件,建立双抗体夹心的ELISA检测方法；同时评价所建立方法的灵敏度、线性范围、特异性、稳定性等性能指标.结果 所建立方法的灵敏度为18.5 ng/ml,线性范围为25～ 500 ng/ml,批内、批间变异均＜10％.检测GP73临床高、中、低值血清回收率为分别为95.3％、92.6％和103.7％；在4℃和37℃条件下分别进行了3、5、7d的稳定性考察,线性相关系数均＞0.98,标准偏差＜10％.结论 成功建立了定量检测血清中GP73的方法,该方法能够满足临床检测的需求.
目的 建立人血清中高爾基體蛋白73(GP73)的定量酶聯免疫吸附試驗(ELISA)檢測方法.方法 辣根過氧化物酶標記GP73單抗,優化各類反應液的工作濃度和各種反應條件,建立雙抗體夾心的ELISA檢測方法；同時評價所建立方法的靈敏度、線性範圍、特異性、穩定性等性能指標.結果 所建立方法的靈敏度為18.5 ng/ml,線性範圍為25～ 500 ng/ml,批內、批間變異均＜10％.檢測GP73臨床高、中、低值血清迴收率為分彆為95.3％、92.6％和103.7％；在4℃和37℃條件下分彆進行瞭3、5、7d的穩定性攷察,線性相關繫數均＞0.98,標準偏差＜10％.結論 成功建立瞭定量檢測血清中GP73的方法,該方法能夠滿足臨床檢測的需求.
목적 건립인혈청중고이기체단백73(GP73)적정량매련면역흡부시험(ELISA)검측방법.방법 랄근과양화물매표기GP73단항,우화각류반응액적공작농도화각충반응조건,건립쌍항체협심적ELISA검측방법；동시평개소건립방법적령민도、선성범위、특이성、은정성등성능지표.결과 소건립방법적령민도위18.5 ng/ml,선성범위위25～ 500 ng/ml,비내、비간변이균＜10％.검측GP73림상고、중、저치혈청회수솔위분별위95.3％、92.6％화103.7％；재4℃화37℃조건하분별진행료3、5、7d적은정성고찰,선성상관계수균＞0.98,표준편차＜10％.결론 성공건립료정량검측혈청중GP73적방법,해방법능구만족림상검측적수구.
Objective To establish enzyme-linked immunosorbent assay (ELISA) for quantitative detection of Golgi protein73 (GP73) in serum.Methods A sandwich reaction was preformed with horseradish peroxidase labeled monoclonal antibody of GP73 as the catalytic enzyme.Several reactions liquid's concentration and reaction conditions were optimized.The method was evaluated in all aspects such as linear range,sensitivity,specificity,stability and so on.Results The linear range was 25-500 ng/ml.The detection limit was 18.5 ng/ml.Inter-assay and intra-assay RSD were both less than 10％.The recoveries of three different spiked concentration samples were 95.3％,92.6％ and 103.7％.After stored at 4℃ and 37℃ for 3,5,7 days,the analysis showed correlation coefficient higher than 0.98 and RSD lower than 10％.Conclusion Established ELISA for quantity determination of serum GP73 has high accuracy,sensitivity and repeatability.
