Chan Cangel P Y, Sum Ka Wai, Cheung Kwan Yee, Glatz Jan F C, Sanderson John E, Hempel Albrecht, Lehmann Matthias, Renneberg Ilka, Renneberg Reinhard
Department of Chemistry, Hong Kong University of Science and Technology, Hong Kong SAR, Kowloon, PR China.
J Immunol Methods. 2003 Aug;279(1-2):91-100. doi: 10.1016/s0022-1759(03)00243-6.
Using human heart-type fatty acid-binding protein (H-FABP) as an early cardiac marker to confirm or exclude a diagnosis of acute myocardial infarction (AMI) soon after admission requires a rapid assay system. Due to the requirement of skillful technicians and complicated assay procedures, most immunochemical assays for H-FABP are of limited use for routine clinical practice. In the present study, we describe a rapid lateral-flow assay for detection of H-FABP. Fifty-one human samples were evaluated using a conventional ELISA and a newly developed lateral-flow assay. A good agreement between the two methods was found according to Bland and Altman plot. The correlation found was y=0.9685 x -0.6270 (r(2)=0.9585). The detector antibody labeled with colloidal gold was mixed with those without label to extend the linear range of the calibration curve up to 125 microg/l H-FABP with r(2)=0.9832. The detection limit of the assay was 2.8 microg/l. The test-strips can be stored either at 4 degrees C and room temperature for up to 1 year without significant loss of activity. Finally, a one-step FABP test so-called CardioDetect(R), which was derived from the serum lateral-flow assay has been designed for qualitative determination of H-FABP in whole blood samples. It requires no sample pretreatment and gives results within 15 min. Thirty-eight patients presenting with chest pain and suspected AMI were studied. Using an upper reference level of 7 microg/l, the specificity of the rapid test was 94%. Both sensitivity and negative predictive value (NPV) were 100%, implying that 100% of non-AMI patients could be excluded with no false-negative results. With this rapid and sensitive immunotest, H-FABP could soon be introduced into clinical practice.
入院后不久，使用人心型脂肪酸结合蛋白（H-FABP）作为早期心脏标志物来确诊或排除急性心肌梗死（AMI）需要一个快速检测系统。由于对技术人员技能要求高且检测程序复杂，大多数针对H-FABP的免疫化学检测在常规临床实践中的应用有限。在本研究中，我们描述了一种用于检测H-FABP的快速侧向流动检测法。使用传统酶联免疫吸附测定（ELISA）和新开发的侧向流动检测法对51份人类样本进行了评估。根据布兰德-奥特曼图，发现两种方法之间具有良好的一致性。发现的相关性为y = 0.9685x - 0.6270（r² = 0.9585）。将胶体金标记的检测抗体与未标记的抗体混合，以将校准曲线的线性范围扩展至高达125μg/l的H-FABP，r² = 0.9832。该检测法的检测限为2.8μg/l。试纸条可在4℃和室温下保存长达1年，活性无明显损失。最后，设计了一种源自血清侧向流动检测法的一步法FABP检测法，即CardioDetect®，用于定性测定全血样本中的H-FABP。它无需样品预处理，15分钟内即可得出结果。对38例出现胸痛并疑似AMI的患者进行了研究。使用7μg/l的上限参考值，快速检测法的特异性为94%。灵敏度和阴性预测值（NPV）均为100%，这意味着100%的非AMI患者可以被排除，且无假阴性结果。通过这种快速且灵敏的免疫检测法，H-FABP很快就能应用于临床实践。
