Autoimmune retinopathy laboratory findings: Difference between revisions
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==Laboratory Findings== | ==Laboratory Findings== | ||
Lab techniques that may aid in the diagnosis of autoimmune retinopathy include immunohistochemistry (IHC), Western blot, and enzyme-linked immunosorbent assay (ELISA). | Lab techniques that may aid in the diagnosis of autoimmune retinopathy include [[Immunohistochemistry|immunohistochemistry (IHC)]], [[Western blot]], and [[ELISA test|enzyme-linked immunosorbent assay (ELISA)]]. | ||
These tests may detect the circulating antirational antibodies, with each having their own advantages and disadvantages. | These tests may detect the circulating antirational antibodies, with each having their own advantages and disadvantages. | ||
=== 1) Immunohistochemistry (IHC) === | ===1) Immunohistochemistry (IHC)=== | ||
In IHC, patient’s serum is incubated with section of normal retina. A secondary antibody is then used to detect the binding of the patient serum to normal retina section. Once disadvantage of this method is that, it is able to detect the total antirational antibody level in the patient’s serum but it can’t detect antibodies against specific proteins. | In IHC, patient’s serum is incubated with section of normal retina. A secondary antibody is then used to detect the binding of the patient serum to normal retina section. Once disadvantage of this method is that, it is able to detect the total antirational antibody level in the patient’s serum but it can’t detect antibodies against specific proteins. | ||
=== 2) Western blot === | ===2) Western blot=== | ||
In Western blot, electrophoresis is used to separate retinal extract or pure retinal proteins on a membrane. The membrane is incubated with patient’s serum and the binding of the patient’s serum to the proteins on the membrane is detected by using a secondary antibody. When a band is evident on a Western blot against the retinal extract suggesting antibodies against a specific protein, then a confirmatory blot needs to be performed using that specific protein. | In Western blot, electrophoresis is used to separate retinal extract or pure retinal proteins on a membrane. The membrane is incubated with patient’s serum and the binding of the patient’s serum to the proteins on the membrane is detected by using a secondary antibody. When a band is evident on a Western blot against the retinal extract suggesting antibodies against a specific protein, then a confirmatory blot needs to be performed using that specific protein. | ||
=== 3) ELISA === | ===3) ELISA=== | ||
ELISA involves small wells that are coated with a specific protein, then patient’s serum is added followed by a secondary antibody, which detects binding. It is similar to IHC and Western blot. <ref name="pmid25730621">{{cite journal| author=Forooghian F, Cao S, Cui J, Matsubara JA| title=The enigma of autoimmune retinopathy. | journal=Int Ophthalmol Clin | year= 2015 | volume= 55 | issue= 2 | pages= 81-91 | pmid=25730621 | doi=10.1097/IIO.0000000000000063 | pmc=4942272 | url=https://www.ncbi.nlm.nih.gov/entrez/eutils/elink.fcgi?dbfrom=pubmed&tool=sumsearch.org/cite&retmode=ref&cmd=prlinks&id=25730621 }} </ref> | ELISA involves small wells that are coated with a specific protein, then patient’s serum is added followed by a secondary antibody, which detects binding. It is similar to IHC and Western blot. <ref name="pmid25730621">{{cite journal| author=Forooghian F, Cao S, Cui J, Matsubara JA| title=The enigma of autoimmune retinopathy. | journal=Int Ophthalmol Clin | year= 2015 | volume= 55 | issue= 2 | pages= 81-91 | pmid=25730621 | doi=10.1097/IIO.0000000000000063 | pmc=4942272 | url=https://www.ncbi.nlm.nih.gov/entrez/eutils/elink.fcgi?dbfrom=pubmed&tool=sumsearch.org/cite&retmode=ref&cmd=prlinks&id=25730621 }} </ref> | ||
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Editor-In-Chief: C. Michael Gibson, M.S., M.D. [1]; Associate Editor(s)-In-Chief: M. Hassan, M.B.B.S
Overview
Lab techniques that may aid in the diagnosis of autoimmune retinopathy include immunohistochemistry (IHC), Western blot, and enzyme-linked immunosorbent assay (ELISA).
Laboratory Findings
Lab techniques that may aid in the diagnosis of autoimmune retinopathy include immunohistochemistry (IHC), Western blot, and enzyme-linked immunosorbent assay (ELISA).
These tests may detect the circulating antirational antibodies, with each having their own advantages and disadvantages.
1) Immunohistochemistry (IHC)
In IHC, patient’s serum is incubated with section of normal retina. A secondary antibody is then used to detect the binding of the patient serum to normal retina section. Once disadvantage of this method is that, it is able to detect the total antirational antibody level in the patient’s serum but it can’t detect antibodies against specific proteins.
2) Western blot
In Western blot, electrophoresis is used to separate retinal extract or pure retinal proteins on a membrane. The membrane is incubated with patient’s serum and the binding of the patient’s serum to the proteins on the membrane is detected by using a secondary antibody. When a band is evident on a Western blot against the retinal extract suggesting antibodies against a specific protein, then a confirmatory blot needs to be performed using that specific protein.
3) ELISA
ELISA involves small wells that are coated with a specific protein, then patient’s serum is added followed by a secondary antibody, which detects binding. It is similar to IHC and Western blot. [1]
References
- ↑ Forooghian F, Cao S, Cui J, Matsubara JA (2015). "The enigma of autoimmune retinopathy". Int Ophthalmol Clin. 55 (2): 81–91. doi:10.1097/IIO.0000000000000063. PMC 4942272. PMID 25730621.