Primary culture Schwann cell IgG and IgM ICC is performed to test Zika-GBS patients staining pattern.
Samples: 11 sera from Zika-GBS (Willison cohort), and a positive and a negative control.
Protocol:
- Primary culture Schwann cells grown in coverslips were fixed with PFA 4 % 10 min, blocked with Goat 5% in PBS 1h, and frozen at -80ºC.
- Thaw these cells with a little Goat 5% in PBS
- Patient’s sera 1/40 for IgM and 1/100 for IgG 1h at RT
- Wash 3 times with PBS.
- Secondary Ab GAH 488 IgG or IgM (1/1000) 1h at RT
- Wash 3 times with PBS.
- Mount with Vectashield mounting medium.
Results:
ID (Zika Willison) IgG/IgM RESULT
003 IgG +++
004 IgG ++
005 IgG -
006 IgG -
008 IgG -
010 IgG -
011 IgG -
013 IgG -
021 IgG ++
024 IgG -
025 IgG -
Positive control IgG -
Negative control IgG -
003 IgM +++
004 IgM ++
005 IgM +
006 IgM -
008 IgM -
010 IgM +
011 IgM ++
013 IgM +
021 IgM +
024 IgM ++
025 IgM +
Positive control IgM ++
Negative control IgM -
