Primary culture Schwann cell IgG and IgM ICC is performed to test controls staining pattern.
Samples: 12 sera from controls, and 2 positive controls (Zika-GBS patients)
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:
Sample Result IgG Result IgM
Healthy control (woman 63y) + +
Healthy control (woman 50y) - +
Healthy control (woman 68y) + +
Healthy control (man 69y) + -
Healthy control (woman 57y) + -
Healthy control (man 50y) - +
Healthy control (man 80y) - -
Healthy control (man 28y) - +
Healthy control (woman 69y) - -
Healthy control (man 29y) - +
Healthy control (woman 22y) - +
Healthy control (man 49y) - -
Zika-GBS 005 - +
Zika-GBS 003 ++ ++
