Development of a high-throughput method for screening the dopamine D2 (DRD2) receptor gene polymorphisms based on the LightCycler system.
BACKGROUND
The human dopamine receptor D2 (DRD2) is known to be involved in several mental disorders. A series of studies revealed that the polymorphisms *TaqIA and *C957T are linked to these disorders and might influence drug response.


METHODS
Screening methods were established for the *TaqIA and *C957T polymorphisms. In the past, the technique of choice has been the time and resource consuming PCR with restriction fragment length polymorphism (PCR-RFLP). It served as the reference method for this investigation.


RESULTS
A protocol was successfully developed for detecting *C957T and *TaqIA on the Roche LightCycler system. In addition, an allele frequency study was conducted comparing 98 samples from healthy and 86 samples from mentally disordered subjects for the *C957T and *TaqIA polymorphisms. Allele frequencies in the healthy population turned out to be 50% for the *C957T and 18% for the *TaqIA polymorphism. In the mentally disordered group, allele frequencies occurred with 54% for the *C957T and with 21% for the *TaqIA polymorphism.


CONCLUSIONS
The protocol developed for identifying the *C957T and *TaqIA polymorphisms is suitable for high-throughput screening. Findings of the allele frequency study did not reveal any significant difference in the prevalence of the investigated polymorphisms.
Stefan
Prause
P
PharmGenomics GmbH, Mainz, Germany. stefan.prause@pharmgenomics.com
Moritz
Eidens
E
Alexander
Weise
W
Weber
Matthias
M
Norbert
Dahmen
D
Annette
Wunsch
W
Thomas
Forst
F
Andreas
Pfützner
P
D005787:Gene Frequency; D020022:Genetic Predisposition to Disease; D057166:High-Throughput Screening Assays; D006801:Humans; D008403:Mass Screening / Q000379:methods*; D001523:Mental Disorders / Q000175:diagnosis / Q000235:genetics; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D017448:Receptors, Dopamine D2 / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*
D017448:Receptors, Dopamine D2