CAJ | 학술논문

目的建立一种新的高灵敏度、宽范围的可溶性细胞间黏附分子-1(Sicam-1)生物素-链亲和素(BSA)系统时间分辨荧光免疫分析(TRFIA)方法并用于临床.方法使用2株匹配的单克隆抗体分别作为捕获抗体和检测抗体,利用制备的铕标记链亲和素(SA-Eu3+)作为示踪物并与生物素化的检测抗体特异结合,建立双位点多层夹心法BSA-TRFIA并进行质量控制,考核其灵敏度、特异性、测量范围及方法稳定性等.检测32例血清样品中Sicam-1含量,并与ELISA所测结果进行相关分析.测定31名健康对照者和22例肺部疾病患者血样中的Sicam-1含量.结果该法测定slCAM-1的灵敏度为1.32μL,批内和批间CV分别为4.24%和6.83%,平均回收率为99.87%,线性范围为(2.5～1933)μg/L.该方法测定值与ELISA所测结果高度相关(R2=0.939).与肿瘤坏死因子(TNF)-α、白细胞介素(IL)-8、IL-1、IL-2和IL-6等无交叉反应.溶血、脂血和高胆红素等血清标本对该方法基本无干扰.31名健康对照者测定结果为(365.74±45.03)μg/L.22例肺部疾病患者血样结果示,在炎症和肺损伤的情况下,可见Sicam-1有升高的趋势.结论该Sicam-1 BSA-TRFIA法灵敏度高,可测范围广,可以方便地应用于临床.
목적 건립일충신적고령민도、관범위적가용성세포간점부분자-1(Sicam-1)생물소-련친화소(BSA)계통시간분변형광면역분석(TRFIA)방법병용우림상.방법 사용2주필배적단극륭항체분별작위포획항체화검측항체,이용제비적유표기련친화소(SA-Eu3+)작위시종물병여생물소화적검측항체특이결합,건립쌍위점다층협심법BSA-TRFIA병진행질량공제,고핵기령민도、특이성、측량범위급방법은정성등.검측32례혈청양품중Sicam-1함량,병여ELISA소측결과진행상관분석.측정31명건강대조자화22례폐부질병환자혈양중적Sicam-1함량.결과 해법측정slCAM-1적령민도위1.32μL,비내화비간CV분별위4.24%화6.83%,평균회수솔위99.87%,선성범위위(2.5～1933)μg/L.해방법측정치여ELISA소측결과고도상관(R2=0.939).여종류배사인자(TNF)-α、백세포개소(IL)-8、IL-1、IL-2화IL-6등무교차반응.용혈、지혈화고담홍소등혈청표본대해방법기본무간우.31명건강대조자측정결과위(365.74±45.03)μg/L.22례폐부질병환자혈양결과시,재염증화폐손상적정황하,가견Sicam-1유승고적추세.결론 해Sicam-1 BSA-TRFIA법령민도고,가측범위엄,가이방편지응용우림상.
Objective The soluble intercellular adhesion molecule-1 (sICAM-1) is an effective marker for inflammation. The purpose of this study was to establish a novel, highly sensitive, wide range and time-resolved fluoroimmunoassay for sICAM-1-biotin-streptavidin complex for use in clinical practice.Methods Two coupled monoclonal antibodies were used as capture and detection antibodies. A two-sided multi "sandwich" immunoassay biotin-streptavidin (BSA), time-resolved fluoroimmunoassay (TRFIA) was developed. In conjunction with pre-synthesized SA-Eu3+ specifically bound to biotin detection antibody, it was used as a marker substance for detection of sICAM-1 level in plasma. The sensitivity, specificity, reliability and range of detection were evaluated. The serum slCAM-1 concentrations were assayed in 32 samplesby TRFIA and the results were compared with those by ELISA using correlation analysis. The serum sICAM-1 concentrations in samples of 31 healthy volunteers and 22 patients with lung disease were determined. Results The lower detection limit was 1.32 μg/L. The inter- and intra-assay CVs were 4.24% and 6.83%, respectively. The mean recovery rate was 99.87%. The linear range of the method for slCAM-1 detection was from 2.5 to 1933 μg/L. The results measured by the TRFIA and ELISA methods were closely correlated (R2 = 0.939). There were no cross-reaction with the tumor necrosis factor (TNF)-α, interleukin (IL)-8,IL-1, IL-2 and IL-6. This method was not affected by serum factors such as hemolysis, fat and high blood serum bilirubin levels. The serum slCAM-1 concentration in the 31 health volunteers was (365.74±45.03) μg/L The serum slCAM-1 concentration tended to be higher in patients with inflammatory lung disease or alveolar damage. Conclusion The TRFIA of slCAM-1 has a high sensitivity and a wide detection range, making it feasible and convenient for clinical applications.