Detection of urine neural thread protein for diagnosis of Alzheimer disease and its clinical significance
10.3760/cma.j.issn.1009-9158.2010.01.011
- VernacularTitle:阿尔茨海默病尿神经丝蛋白检测方法的建立及其临床意义
- Author:
Rong WANG
;
Zhijuan JI
;
Shuli SHENG
;
Jianan ZHU
;
Zhiwei ZHAO
;
Ziqing CAO
;
Peichang WANG
;
Xianghong MENG
;
Jingyan ZHANG
- Publication Type:Journal Article
- Keywords:
Alzheimer diseases;
Urinalysis;
Nerve tissue proteins;
Enzyme-linked immunosorbent assay
- From:
Chinese Journal of Laboratory Medicine
2010;33(1):46-50
- CountryChina
- Language:Chinese
-
Abstract:
Objective To develop urine AD7C-NTP diagnostic kit,analyze and evaluate its application value on AD.Methods Immunogenicity AD7C-NTP peptide fragments had synthesized by solidphase methods.The animals immunized to prepare antibodies.After matching screening.mouse antibody was uesed as coating antibody.biotin-labeled rabbit antibody wag used as testing antibody,and horseradish peroxidase was labeied with avidin.The urine AD7C-NTP ELISA detective method was established.The AD7C-NTP levels in morning urine samples of 121 AD patients and 118 age-matched controls were collected.Results AD7C-NTP antibodies were identified.Mouse anti-AD7C-NTP antibody titer in ELISAwas 1:8 000,and rabbit anti-AD7C-NTP antibody titer in ELISA was 1:32 000:WB was uesd to detect human brain specimens and there was a single band with molecular weight of 41 000.The lowst detection limit of ELISA methodology was 0.5μg/L The linear range was 0-10μg/L,normal reference value ≤1.5μg/J,the average recovery rate was 100.2%.The intra and inter of CV were 3.8%,4.5%,7.6%,6.8% respectively.The AD7C-NTP levels[2.25(0.43-8.62)μg/L]of urine in AD group was higher than those in contorl group[0.82(0.47-2.77)μg/L,P<0.01].The positive rates in AD group and control group were 89.3% and 15.3% respectively.The sensitivity Was 89.3%and specificity was 84.7%.Conclusions The animals are immunized with the self-designed synthetic peptide fragment to prepare AD7C-NTP antibodies successfully.The established ELISA method for detection of urine AD7C-NTP with high sensitivity,and precision can be used as an assistant examination in clinical diagnosis of AD.
