Development of a simple multiplex polymerase chain reaction for the simultaneous detection of factor V Leiden and prothrombin 20210A mutations.
BACKGROUND
The demand for thrombophilia testing at the molecular level is increasing, and consequently, the work load of the routine molecular laboratory is also increasing. Efforts to lighten the work load, economize on time, and strive for reduced costs while still maintaining quality assurance are thus necessary.


RESULTS
A multiplex polymerase chain reaction (PCR) for the detection of factor V Leiden and prothrombin 20210A mutations was designed that enables the use of the same inexpensive restriction enzyme, controls for the digestion, and produces easily interpretable results.


CONCLUSIONS
The use of this new multiplex PCR and digestion analysis enabled us to simultaneously perform a routine screen for factor V Leiden and prothrombin 20210A mutations.
Dubreuil Lastrucci
R M
RM
Department of Haematology, School of Pathology, University of the Witwatersrand, Johannesburg, South Africa.
Dawson
D A
DA
Bowden
J H
JH
Marion Münster
M
M
D020413:3' Untranslated Regions / Q000235:genetics*; D000483:Alleles; D004252:DNA Mutational Analysis / Q000379:methods*; D005165:Factor V / Q000235:genetics*; D005166:Factor V Deficiency / Q000175:diagnosis* / Q000235:genetics; D020022:Genetic Predisposition to Disease; D005820:Genetic Testing / Q000379:methods*; D005838:Genotype; D006801:Humans; D017354:Point Mutation; D016133:Polymerase Chain Reaction / Q000379:methods*; D011516:Prothrombin / Q000235:genetics*; D012307:Risk Factors; D019851:Thrombophilia / Q000175:diagnosis / Q000453:epidemiology / Q000235:genetics*
D020413:3' Untranslated Regions; C095381:factor V Leiden; D005165:Factor V; D011516:Prothrombin