Objective: To test IgG positivity towards NF155 in GBS samples from the spain cohort of IGOS + confirmation of positivity of sample 18-860.
SAMPLES: 0-42, 18-860, CTRL – (1 to 5) and CTRL + (LAC)
MATERIALS:
PROCEDURE:
- Coat ELISA plate with NF155 protein (1ugr/mL) in carbonate/bicarbonate buffer (pH 9.6) and incubate O/N at 4ºC rocking.
- The day after, wash plate 3xPBS-Tween 0.1% and block with milk 5% in PBS-Tween for 1 hour, 200uL per well).
- Add patients sera diluted 1/100 (200uL/well) and incubate 1h at RT.
- Wash 3xPBS-Tween 0.1%
- Add secondary antibody using RAH IgG 1/5000 diluted in milk 5% in PBS-Tween and incubate 1h at RT.
- Wash 3xPBS-Tween
- Add TMB solution 10′
- Stop reaction with H2SO4
- Lecture at 450-620 nm.
RESULTS:
The figure of ELISA results is here.
All samples were negative except 18-860.