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In vitro validation of the tear matrix metalloproteinase 9 in-situ immunoassay

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Author(s)
Seung Pil BangMyeong Jin SonHarim KimYou Hyun LeeJong Hwa Jun
Keimyung Author(s)
Lee, You HyunJun, Jong Hwa
Department
Dept. of Ophthalmology (안과학)
Journal Title
Scientific Reports
Issued Date
2020
Volume
10
Issue
1
Abstract
We aimed to validate a tear MMP-9 in-situ immunoassay (InflammaDry) and to identify factors that could affect results or interpretation. Three factors were examined: sample concentration, volume, and time. Recombinant human (rh) MMP-9 (10 or 20 μl; 0, 12.5, 25, 50, 100, 200, 500, and 1,000 ng/ml) was applied to the kit and the detection limit and assay reproducibility were examined. At a rhMMP-9 volume of 10 μl (≥ 50 ng/ml), all positive results were identified by densitometry at 10 and 20 min; however, after 20 min, more than half of the nine ophthalmologists interpreted a positive result. At a rhMMP-9 volume of 20 μl (≥ 25 ng/ml), ophthalmologists and densitometry identified almost all test lines at 10 and 20 min. At 10 μl, densitometry showed a linear dose-response pattern. At 20 μl, densitometry showed a linear dose-response pattern at concentrations up to 500 ng/ml; however, full saturation was achieved at concentrations ≥ 500 ng/ml. When the same amount of rhMMP-9 was applied, the density result increased significantly upon doubling of the solvent volume (i.e., by adding the same volume of PBS to a sample). InflammaDry showed a high inter- and intra-assay coefficient of variation at 10 min (28.4% and 24.7%, respectively). The results of the MMP-9 in-situ immunoassay varied significantly depending on sample volume. Therefore, when interpreting the results, careful attention must be paid to tear volume.
Keimyung Author(s)(Kor)
이유현
전종화
Publisher
School of Medicine (의과대학)
Citation
Seung Pil Bang et al. (2020). In vitro validation of the tear matrix metalloproteinase 9 in-situ immunoassay. Scientific Reports, 10(1), 15126. doi: 10.1038/s41598-020-71977-w
Type
Article
ISSN
2045-2322
Source
https://www.nature.com/articles/s41598-020-71977-w
DOI
10.1038/s41598-020-71977-w
URI
https://kumel.medlib.dsmc.or.kr/handle/2015.oak/43299
Appears in Collections:
1. School of Medicine (의과대학) > Dept. of Ophthalmology (안과학)
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