Detection of soluble HLA class I antigens by ELISA and determination of its reference value in population of Guangdong province.
OBJECTIVE
To explore a method for quantitative detection of soluble human leukocyte antigen class I(HLA-I) antigens, with the assistance of which we attempt to define the reference value of the antigen in the population of Guangdong Province.


METHODS
Sandwich enzyme-linked immunosorbent assay (ELISA) was employed with W6/32 monoclonal antibody serving as the solid phase antibody and beta2 microglobulin antibody as the first antibody. HRP-labeled second antibody and the substrate were added for enzyme-catalyzed coloring. On the basis of the standard curve obtained by sHLA-I standard reagent in serial dilution, the concentration of sHLA-I antigens in the samples of 60 normal subjects from the population of Guangdong province was detected.


RESULTS
The sensitivity of this assay was 2.84 ng/ml with variation coefficients of 5.80% within the same batch and 9.00% between batches. The recovery rate ranged from 98.57% to 100.15%. The level of sHLA-I antigens in normal individuals was 699.54+/-360.10 ng/ml.


CONCLUSIONS
Sandwich ELISA is sensitive, specific and stable in the detection of sHLA-I.
Wu
Tao
T
Department of Transfusion, Nanfang Hospital, First Military Medical University, Guangzhou 510515, China.
Lan
Jiong-Cai
JC
Zhang
Hong
H
Wang
Cong-Rong
CR
Mao
Gao-Cai
GC
D000328:Adult; D002681:China; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000592:standards; D005260:Female; D015395:Histocompatibility Antigens Class I / Q000097:blood*; D006801:Humans; D008297:Male; D008875:Middle Aged; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012995:Solubility
D015395:Histocompatibility Antigens Class I