The New Establishment of Quantitative Determination Method for Food Intolerance Specific IgG
10.3969/j.issn.0253-9896.2013.11.001
- VernacularTitle:新型食物不耐受sIgG定量检测方法的构建
- Author:
Ping XIANG
;
Jing SUN
;
Yujie ZHOU
;
Jie REN
;
Tangyuheng LIU
;
Huiqiang LI
- Publication Type:Journal Article
- Keywords:
food intolerance;
specific IgG;
enzyme-linked immunosorbent assay;
biotin-streptavidin;
egg
- From:
Tianjin Medical Journal
2013;(11):1041-1044
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore a novel quantitative detection method for the concentration of specific IgG (sIgG) in food intolerance, taking egg sIgG detection for the example. Methods A total of 173 patients underwent food allergen sIgG detection were included in this study, and 78 healthy subjects were used as negative controls. The microtiter plates were coated with biotinylated bovine serum albumin (BSA) and linked with streptavidin. Then, the biotinylated egg antigen and specimen were successively added into the wells of plate.After washing, enzyme labeled anti-human IgG was added to establish an antigen indirectly coated liquid-phase reaction patterns of ELISA method. The concentration of the biotinylated allergens and enzyme labeled antibody were optimized and the reaction conditions were determined. This method was used to detect the sIgG in serum samples. Results The biotinylated egg white was selected as antigen, the optimal dilution rate was 1∶2 000,and the most suitable enzyme labeled antibody dilution ratio was 1∶12 000 in this method. The within-run and the between-run coefficients of variation were 4.83%-8.55%and 4.88%-7.93%respectively. The specificity is preferable, the species-crossed reaction rates with crab, cow milk and goat milk were＜10%. There was a good correlation between the assay developed in this study and the food intolerance detection kit provided by United States BIOMERICA Inc ( =0.977X+8.45, r=0.961, P＜0.05). Conclusion The detection method can be used to detect serum sIgG for egg intolerance patients with easy operation, highly accuracy and specificity. Furthermore, it showed the capacity of excellent repeatability and flexibility potential, which provided a good foundation for developing a kind of“personalized”random combination of food varieties ELISA kit.
