[Genetic diagnosis of autosomal dominant polycystic kidney disease using multiplex-PCR].
BACKGROUND
Autosomal Dominant Polycystic Kidney Disease (ADPKD) is the most common life-threatening hereditary disease. Molecular analysis with highly polymorphic short tandem repeats, located in the vicinity of the two genes responsible for the disease (PKD1 and PKD2), is used to confirm diagnosis and give genetic counseling to members of affected families.


METHODS
We have developed a new assay to genotype five PKD1 and four PKD2 markers, based on two multiplex PCR reactions, and capillary electrophoresis analysis. A total of 110 subjects, belonging to 14 affected families, were genotyped to confirm the concordance with the singleplex method used previously.


RESULTS
The amplicons ranged from 95 to 154 bp in length, and complete STR profiles were obtained from 1-5 ng DNA. The specificity of the multiplex PCR system was 88,5% (95%CI= 75,9-95,2), and the sensitivity, 87,9 (95%CI= 76,1-94,6).


CONCLUSIONS
This is a useful strategy that, together with automated computer-based allele detection, allows reliable, simple, faster, and cheaper genetic analysis than the previous singleplex method.
Buset Ríos
N
N
Unidad de Investigación, Las Palmas de Gran Canaria.
Rodríguez Pérez
J C
JC
Sánchez
J J
JJ
Hernández
C R
CR
Hernández
E
E
Torres Galván
M J
MJ
D005260:Female; D006801:Humans; D008297:Male; D018895:Microsatellite Repeats; D016891:Polycystic Kidney, Autosomal Dominant / Q000175:diagnosis* / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D050396:TRPP Cation Channels / Q000032:analysis / Q000235:genetics
D050396:TRPP Cation Channels