| Title |
Development of a highly sensitive and reliable enzyme-linked immunosorbent assay for MUC5AC in human tears extracted from Schirmer strips
|
|---|---|
| Published in |
Clinical Ophthalmology, August 2018
|
| DOI | 10.2147/opth.s170552 |
| Pubmed ID | |
| Authors |
Hideki Miyake, Naoto Mori, Hidetoshi Mano, Takahiro Imanaka, Masatsugu Nakamura |
| Abstract |
Reliable measurement of MUC5AC in human tears is essential for elucidation of the pathophysiological role of MUC5AC in dry eye disease. The purpose of this study was to develop a sensitive and reliable method for measurement of MUC5AC in human tear samples extracted from Schirmer strips by modifying a commercially available ELISA. MUC5AC was extracted from Schirmer strips containing human tears by PBS with various concentrations of polysorbate 20. The extracts were treated with neuraminidase A to cleave the sialic acids in MUC5AC. An ELISA plate was blocked to prevent nonspecific binding. The rate of extraction of MUC5AC from Schirmer strips, linearity of dilution, limit of quantification, calibration range, and intra-assay and inter-assay reproducibility were examined. MUC5AC was extracted using polysorbate 20 in a concentration-dependent manner. Extraction was more efficient at 37°C than at 25°C. The signal-to-noise ratio of the assay was dramatically increased by treatment with neuraminidase A. Treatment with a blocking reagent before incubation produced good linearity of dilution. The inter-assay and intra-assay coefficients of variation were ≤16.6%. The relative error was within 13%. We developed an efficient method for extraction of MUC5AC from Schirmer strips and a highly sensitive, reliable assay for MUC5AC in human tear samples using a commercially available ELISA kit. This method will aid in our understanding of the pathophysiology of dry eye, assessment of the effects of treatment in daily practice, and selection of appropriate therapeutic agents for patients. |
X Demographics
The data shown below were collected from the profile of 1 X user who shared this research output. Click here to find out more about how the information was compiled.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 1 | 100% |
Demographic breakdown
| Type | Count | As % |
|---|---|---|
| Members of the public | 1 | 100% |
Mendeley readers
The data shown below were compiled from readership statistics for 15 Mendeley readers of this research output. Click here to see the associated Mendeley record.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 15 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Ph. D. Student | 3 | 20% |
| Student > Doctoral Student | 2 | 13% |
| Other | 1 | 7% |
| Professor > Associate Professor | 1 | 7% |
| Student > Postgraduate | 1 | 7% |
| Other | 0 | 0% |
| Unknown | 7 | 47% |
| Readers by discipline | Count | As % |
|---|---|---|
| Medicine and Dentistry | 2 | 13% |
| Veterinary Science and Veterinary Medicine | 1 | 7% |
| Pharmacology, Toxicology and Pharmaceutical Science | 1 | 7% |
| Nursing and Health Professions | 1 | 7% |
| Biochemistry, Genetics and Molecular Biology | 1 | 7% |
| Other | 2 | 13% |
| Unknown | 7 | 47% |
Attention Score in Context
This research output has an Altmetric Attention Score of 1. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 15 September 2018.
All research outputs
#22,767,715
of 25,385,509 outputs
Outputs from Clinical Ophthalmology
#3,207
of 3,714 outputs
Outputs of similar age
#299,007
of 341,886 outputs
Outputs of similar age from Clinical Ophthalmology
#24
of 30 outputs
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