Microplate chemiluminescence enzyme immunoassay for the quantitative analysis of tissue inhibitor of metalloproteinases I / 中华实验和临床病毒学杂志
Chinese Journal of Experimental and Clinical Virology
;
(6): 298-300, 2013.
Article
in Chinese
| WPRIM
| ID: wpr-318036
ABSTRACT
<p><b>OBJECTIVE</b>To establish microplate chemiluminescence enzyme immunoassay (CLEIA) for quantitative analysis of tissue inhibitor of metalloproteinases I (TIMP I) in human serum.</p><p><b>METHODS</b>A sandwich reaction was preformed with horseradish peroxidase(HRP) labeled monoclonal antibody of TIMP I as the catalytic enzyme and the H2O2-luminol as the luminescence reagent. Several physical and chemical parameters were studied and optimized such as immunoreaction conditions, the dilution ratio of TIMP I-HRP, luminescence reaction time and so on. In order to evaluate the method, recovery test, heat stabilization test and comparison test were carried out.</p><p><b>RESULTS</b>The linear range was 0. 2-12 ng/ml with r = 0.996. The detection limit was 0.12 ng/ml. Inter-assay and intra-assay RSD were both less than 10%. The recoveries of three different spiked concentration samples were 100.6%, 96.5% and 106.5%. After stored at 4 degrees C and 37 degrees C for 3, 5, 7 days, the analysis showed correlation coefficient higher than 0. 998 and RSD lower than 6%. The detected results with CLEIA closely corresponded to those with imported ELISA in 60 patients sera with liver fibrosis.</p><p><b>CONCLUSION</b>Established CLEIA for quantity determination of serum TIMP I has high accuracy, sensitivity and repeatability.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Blood
/
Immunoenzyme Techniques
/
Tissue Inhibitor of Metalloproteinase-1
/
Diagnosis
/
Liver Cirrhosis
/
Luminescent Measurements
/
Methods
Type of study:
Diagnostic study
Limits:
Adult
/
Aged
/
Female
/
Humans
/
Male
Language:
Chinese
Journal:
Chinese Journal of Experimental and Clinical Virology
Year:
2013
Type:
Article
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