CAJ | 학술논문

目的建立人血清中Ⅲ型前胶原氨基端肽(PⅢNP)的定量微孔板化学发光酶免疫检测方法.方法辣根过氧化物酶标记PⅢNP单抗,选用鲁米诺化学发光体系检测,优化各类反应液的工作浓度和各种反应条件,建立双抗体夹心的检测方法；同时评价所建立方法的灵敏度、特异性、线性范围、稳定性等性能指标；并应用临床血清与进口试剂进行比对实验.结果所建立方法的线性范围为0.8 ～ 85 ng/ml,灵敏度0.5 ng/ml,批内、批间变异均＜10％.检测PⅢNP的临床高、中、低值血清回收率分别为96.2％、91.2％和101.1％；在4℃和37℃条件下分别进行了3d、5d、7d的稳定性考察,线性相关系数均＞0.99,标准偏差＜6％；比对实验分析显示与进口试剂相关性具有统计学意义.结论成功建立了定量PⅢNP化学发光酶免疫分析方法,并且有较好的准确性、灵敏度、重复性,与进口试剂检测结果等效.
목적 건립인혈청중Ⅲ형전효원안기단태(PⅢNP)적정량미공판화학발광매면역검측방법.방법 랄근과양화물매표기PⅢNP단항,선용로미낙화학발광체계검측,우화각류반응액적공작농도화각충반응조건,건립쌍항체협심적검측방법；동시평개소건립방법적령민도、특이성、선성범위、은정성등성능지표；병응용림상혈청여진구시제진행비대실험.결과 소건립방법적선성범위위0.8 ～ 85 ng/ml,령민도0.5 ng/ml,비내、비간변이균＜10％.검측PⅢNP적림상고、중、저치혈청회수솔분별위96.2％、91.2％화101.1％；재4℃화37℃조건하분별진행료3d、5d、7d적은정성고찰,선성상관계수균＞0.99,표준편차＜6％；비대실험분석현시여진구시제상관성구유통계학의의.결론 성공건립료정량PⅢNP화학발광매면역분석방법,병차유교호적준학성、령민도、중복성,여진구시제검측결과등효.
Objective To establish chemiluminescence enzyme immunoassay (CLEIA) for quantitative detection of procollagen Ⅲ N-terminal peptide (P ⅢNP) in serum.Methods A sandwich reaction was preformed with horseradish peroxidase labeled monoclonal antibody of P Ⅲ NP as the catalytic enzyme and the luminol as the luminescence reagent.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.The CLEIA was compared with imported ELISA kits,by detecting clinical serum.Results The linear range was 0.8-85 ng/ml.The detection limit was 0.5 ng/ml.Inter-assay and intra-assay RSD were both less than 10％.The recoveries of three different spiked concentration samples were 96.2％,91.2％ and 101.1％.After stored at 4℃ and 37℃ for 3,5,7 days,the analysis showed correlation coefficient higher than 0.99 and RSD lower than 6％.The detected results of clinical sera with CLEIA closely corresponded to those with imported ELISA.Conclusion Established CLEIA for quantity determination of serum PⅢ NP has high accuracy,sensitivity and repeatability.