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Prozone phenomenon observed in indirect immunofluorescence assay by antibodies against neuronal antigens
Annamma Mathai1, Suprabha Panicker1, Sudheeran Kannoth1
1Department of Neurology, Amrita Visvavidyapeetham, Amrita University, Amrita Institute of Medical sciences, Kochi 682041, Kerala, India; Neuroimmunology Laboratory, Amrita Visvavidyapeetham, Amrita University, Amrita Institute of Medical sciences, Kochi 682041, Kerala, India.
Abstract:
A marked prozone effect was observed in indirect immunofluorescence with human sera and human cerebrospinal fluid in two clinical cases involving breast carcinoma with paraneoplastic neuronal antibodies, and anti- N-methyl-D-aspartic acid (NMDA) receptor antibodies. Anti-Yo antibodies and anti-NMDA antibodies were not detectable under high concentrations (1:10 serum dilution and neat CSF respectively) but showed a true effect when sufficiently diluted at 1:80 and 1:5 respectively. This paper demonstrates that prozone effects have their occurrences in indirect immunofluorescence, and clinicians and laboratory technicians should be wary of its implications during screening of autoantibody markers in neurological diseases.
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01:12Immunofluorescence Microscopy
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The technique begins with separating proteins from the sample using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), followed by protein transfer, immunoblotting, and finally, protein detection.