FMF StripAssay

The Viennalab FMF StripAssay is a qualitative genetic test for the targeted analysis of 12 frequent mutations in the MEFV gene associated with Familial Mediterranean Fever.

Clinical background

MEFV encodes pyrin, a protein that regulates the inflammatory response. Mutations lead to recurrent episodes of fever, peritonitis, pleuritis and arthritis. The severity varies with the genotype, and so does the risk of systemic AA amyloidosis, in which serum amyloid A fragments are deposited in tissue and cause irreversible organ damage, most often in the kidney. Colchicine as prophylaxis lowers that risk, which is what makes the genotype consequential.

Mutations covered

The strip covers E148Q, P369S, F479L, M680I in two variants (c.2040G>C and c.2040G>A), I692del, M694V, M694I, K695R, V726A and A744S, with the corresponding HGVS notation and rs numbers listed in the IFU. Note that M680I occupies two of the twelve lines, one for each nucleotide change.

Method and sample

Multiplex PCR with biotin labelling is followed by hybridisation on the test strip and detection with streptavidin-alkaline phosphatase and a colour substrate, read by eye or with a scanner and software. Every strip carries a control line for the PCR reaction and a hybridisation control. Fresh or frozen blood with EDTA as anticoagulant is used; the kit contains the lysis solution and GENxTRACT resin, and Taq DNA Polymerase is supplied (75 U). For DNA prepared by another method, 2 to 10 ng/µl is specified.

• Lysis Solution: 50 ml
• GENXTRACT™ Resin: 5 ml
• Amplification Mix: 500 µl (Yellow Cap)
• Taq Dilution Buffer: 500 µl (Transparent Cap)
• DNAT: 1.5 ml (Blue Cap, Warning)
• Typing Trays: 3 included
• Test Strips: 20 included
• Hybridization Buffer: 25 ml (White Cap)
• Wash Solution A: 80 ml (White Cap)
• Conjugate Solution: 25 ml
• Wash Solution B: 80 ml
• Color Developer: 25 ml (Warning)
• Collector™ Sheet: 1 included