CAJ | 학술논문

目的建立人血清透明质酸夹心化学发光免疫检测方法.方法用透明质酸结合蛋白分别进行固相载体包被和与辣根过氧物酶偶联,建立定量测定人血清透明质酸的夹心化学发光免疫检测法,并对该方法进行方法学和临床应用性评价. 结果所建立血清透明质酸化学发光免疫检测法灵敏度为0.66μg/L,线性范围在1.8~1000μg/L,批内变异系数低于7.05%,批间变异系数低于8.0%,平均添加回收率为103.1%,稀释回收率为106.8%,与肝纤维化其他血清标志物 IV 型胶原、III 型前胶原和层黏连蛋白的交叉反应率分别只有0.19%、0.16%和0.17%,样本中添加的抗凝剂及胆红素、胆固醇、甘油三酯等物质对检测结果没有影响.辽宁抚顺市传染病医院用该方法检测737份样本的临床总符合率为98.64%;检测197份样本透明质酸含量的结果与美国 Corgenix 试剂盒检测结果的相关性为 y =0.855x +26.874(r=0.9877). 结论该方法灵敏度高、特性好、线性范围宽,有良好的准确性和重复性,完全可用于临床样本的肝纤维化检测.
목적건립인혈청투명질산협심화학발광면역검측방법.방법용투명질산결합단백분별진행고상재체포피화여랄근과양물매우련,건립정량측정인혈청투명질산적협심화학발광면역검측법,병대해방법진행방법학화림상응용성평개. 결과소건립혈청투명질산화학발광면역검측법령민도위0.66μg/L,선성범위재1.8~1000μg/L,비내변이계수저우7.05%,비간변이계수저우8.0%,평균첨가회수솔위103.1%,희석회수솔위106.8%,여간섬유화기타혈청표지물 IV 형효원、III 형전효원화층점련단백적교차반응솔분별지유0.19%、0.16%화0.17%,양본중첨가적항응제급담홍소、담고순、감유삼지등물질대검측결과몰유영향.료녕무순시전염병의원용해방법검측737빈양본적림상총부합솔위98.64%;검측197빈양본투명질산함량적결과여미국 Corgenix 시제합검측결과적상관성위 y =0.855x +26.874(r=0.9877). 결론해방법령민도고、특성호、선성범위관,유량호적준학성화중복성,완전가용우림상양본적간섬유화검측.
Objective To develop a chemiluminescence immunoassay (CLIA) for the detection of hyaluronic acid (HA) from human sera. @@@@Methods The quantitive CLIA was established based on double-antibody sandwich technology. Hyaluronic acid binding protein (HABP) was coated on microtiter plates and labeled with horseradish perxidase (HRP) respectively. Then the methodological evaluation and clinical application study about the method was assessed. @@@@Results The sensitivity of the CLIA method is 0.66μg/L, and linear range is 1.8~1000μg/L. Coefficients of variation (CV) for within-run and between-run assays is lower than 7.05%and 8.0%, respectively. The addition recovery and dilution recovery rate is 103.1%and 106.8%, respectively. The cross reactivity with Laminin, type IV collagen and type III procollagen is 0.19%, 0.16%and 0.17%, respectively. There is little effect of anticoagulants, bilirubin, cholesterol or triglyceride added in the serum samples on the assay. The conformance rate of 737 clinical serum samples is 98.64%as assayed by the method in Fushun infectious disease hospital. The correlation of HA detection assay in 197 serum samples between the CLIA method and HA Test Kit (Corgenix, USA) is y =0.855x+26.874 (r=0.9877). @@@@Conclusions The developed CLIA for the detection of HA from human sera is a sensitive and specific method with broad linear range, high accuracy and reproducibility. It can be applied in clinic for the detection of liver fibrosis.