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乳胶凝集法和定量法测定小儿血清中C反应蛋白(CRP)的效能

Efficacy of latex agglutination and quantification methods for determination of C-reactive protein (CRP) in pediatric sera.

作者信息

Wadsworth C, Wadsworth E

出版信息

Clin Chim Acta. 1984 Apr 27;138(3):309-18. doi: 10.1016/0009-8981(84)90138-4.

Abstract

Described is a procedure for serum C-reactive protein (CRP) determination, consisting of a semiquantitative rapid CRP latex agglutination test, using dilutions of the serum, and the quantitating spot immunoprecipitate assay (SIA). These methods are performed with standard laboratory equipment using no more than 30 microliters of serum for both assays. With visual inspection, the SIA results are available one to two hours after blood sampling. CRP levels obtained by agglutination testing with five batches of latex reagents coated with rabbit anti-CRP agree well with the quantification, i.e. essentially 100% for the negative sera and about 80% for positive sera containing greater than 40 mg CRP/l. The remaining 20% of the samples are classified as low positive at 10 to 40 mg/l on agglutination. False positive or negative agglutination findings are below 2.3% with concordance at 88% between SIA and the CRP agglutination with rabbit antibodies. Two lots of CRP-latex reagents coated with sheep antibodies, however, gave 15.3% and 10.1% false positive findings and poor concordance with SIA ratings, particularly for low positive sera at only 20 and 29%. SIA is suggested for CRP quantification because it compares well with radial immunodiffusion in accuracy (less than 91%) and provides results in 2 h rather than 1-2 days. Rocket electroimmunoassay is less reliable with lower ratings than found in SIA, probably due to the electrophoretic heterogeneity of CRP. This is demonstrated for two of three purified CRP preparations, for which varying agglutination is seen. The combination of methods is especially recommended for diagnosis and monitoring of CRP in infectious processes in neonates and infants because of the required small sample volume--0.5 ml heel-prick blood--the rapidity of reliable (greater than 80%) reporting and the possibility of rating sera with moderate levels of CRP.

摘要

本文描述了一种血清C反应蛋白(CRP)的检测方法,该方法包括使用血清稀释液的半定量快速CRP乳胶凝集试验和定量斑点免疫沉淀试验(SIA)。这些方法使用标准实验室设备进行,两种检测均只需不超过30微升的血清。通过目视检查,SIA结果在采血后1至2小时即可获得。用五批包被兔抗CRP的乳胶试剂进行凝集试验得到的CRP水平与定量结果吻合良好,即阴性血清的吻合率基本为100%,CRP含量大于40mg/L的阳性血清的吻合率约为80%。其余20%的样本在凝集试验中被归类为低阳性,CRP含量为10至40mg/L。凝集试验的假阳性或假阴性结果低于2.3%,SIA与兔抗体CRP凝集试验的一致性为88%。然而,两批包被羊抗体的CRP乳胶试剂的假阳性率分别为15.3%和10.1%,与SIA评级的一致性较差,尤其是低阳性血清的一致性仅为20%和29%。建议使用SIA进行CRP定量,因为其在准确性方面(小于91%)与放射免疫扩散法相当,且能在2小时而非1至2天内得出结果。火箭免疫电泳法的可靠性较低,评级低于SIA,这可能是由于CRP的电泳异质性所致。对三种纯化的CRP制剂中的两种进行了验证,观察到了不同的凝集情况。由于所需样本量小(足跟穿刺血0.5ml)、可靠报告速度快(大于80%)以及能够对CRP水平中等的血清进行评级,因此特别推荐将这些方法联合用于新生儿和婴儿感染性疾病中CRP的诊断和监测。

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