血清Ⅲ型前胶原N端肽化学发光酶免疫分析定量检测

[Quantitative detection of serum procollagen III N-terminal peptide chemiluminescence enzyme immunoassay].

作者信息

Liu Jia, Feng Yan-Qing, Song Yong-Ji, Hou Jun, Chen Lin, Zhao Jing, Liu Ai-Xia, Guo Jing-Xia, Xu Jun, Wei Hong-Shan, Li Bo-An

出版信息

Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. 2013 Oct;27(5):385-7.

PMID:24645331

Abstract

OBJECTIVE

To establish chemiluminescence enzyme immunoassay (CLEIA) for quantitative detection of procollagen III N-terminal peptide (P III NP) 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 degrees C and 37 degrees C 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 P III NP has high accuracy, sensitivity and repeatability.

摘要

目的

建立化学发光酶免疫分析法（CLEIA）定量检测血清中Ⅲ型前胶原氨基端肽（PⅢNP）。

方法

以辣根过氧化物酶标记的PⅢNP单克隆抗体为催化酶，鲁米诺为发光试剂，进行夹心反应。对几种反应液浓度及反应条件进行优化。从线性范围、灵敏度、特异性、稳定性等方面对该方法进行评价。通过检测临床血清，将CLEIA法与进口ELISA试剂盒进行比较。

结果

线性范围为0.8 - 85 ng/ml。检测限为0.5 ng/ml。批间和批内相对标准偏差均小于10%。3种不同加标浓度样品的回收率分别为96.2%、91.2%和101.1%。在4℃和37℃分别保存3、5、7天后，分析显示相关系数高于0.99，相对标准偏差低于6%。CLEIA法检测临床血清的结果与进口ELISA法的结果密切相符。