A rapid and cost-effective method for analysis of three common genetic risk factors for thrombosis.
A simple, rapid and cost-effective method for the analysis of three of the most widely screened genetic risk factors for thrombosis has been established. The protocol developed uses blood spots stored on filter paper (Guthrie spots) as well as DNA extracted from anticoagulated blood. The use of Guthrie spots taken at birth enables the retrospective study of patients who develop thrombotic complications without necessitating resampling. Following isolation of DNA, conventional fluorescence-labelled polymerase chain reaction (PCR) is performed using a thermostable DNA polymerase. Denatured, single-stranded PCR products are analysed on a semi-automated capillary-based genetic analyser, the data being stored electronically. This sensitive protocol obviates the need for endonuclease digestion and the associated gel running and documentation, and leads to a reduction in the recurrent costs of laboratory consumables.
The Reference Centre for Haemostatic and Thrombotic Disorders, St Thomas' Hospital, London, UK. jacqueline.cutler@gstt.sthames.nhs.uk
J A
JA
Cutler
M J
MJ
Mitchell
K
K
Greenslade
M P
MP
Smith
G F
GF
Savidge
D001800:Blood Specimen Collection / Q000379:methods; D003362:Cost-Benefit Analysis; D004252:DNA Mutational Analysis / Q000379:methods / Q000592:standards; D001330:Electronic Data Processing; D005456:Fluorescent Dyes; D005819:Genetic Markers; D005820:Genetic Testing / Q000191:economics / Q000379:methods* / Q000592:standards; D006801:Humans; D018807:Polymorphism, Single-Stranded Conformational; D012189:Retrospective Studies; D012307:Risk Factors; D013927:Thrombosis / Q000175:diagnosis / Q000235:genetics*
D005456:Fluorescent Dyes; D005819:Genetic Markers