Well-devised quantification analysis for duplication mutation of Duchenne muscular dystrophy aimed at preimplantation genetic diagnosis.
OBJECTIVE
Preimplantation genetic diagnosis (PGD) has been performed for deletion and point mutation type of Duchenne muscular dystrophy (DMD). Our aim was to develop a PGD technique, not yet established, to directly detect duplication mutation instead of substitute diagnosis similar to gender determination.


METHODS
Our method is based on comparative quantification using conventional duplex PCR, real-time PCR and gender determination. We evaluated this method in single lymphocytes from a duplication type of DMD patient and a normal male.


RESULTS
There was a significant difference in the mean values of the ratios (the mutation locus/a normal reference): mean value +/- SE was 1.84 +/- 0.15 in the duplication patient, and 1.00 +/- 0.09 in the normal male (p < 0.001).


CONCLUSIONS
It is suggested that our comparative quantification method could be a new option in PGD for carriers with duplication mutation who wish to have an unaffected son.
Nakabayashi
Akira
A
Department of Obstetrics and Gynecology, Keio University School of Medicine, 35 Shinanomachi Shinjuku-ku, Tokyo 160-8582, Japan. akira-na@kanagawa.saiseikai.or.jp
Sueoka
Kou
K
Tajima
Hiroto
H
Sato
Kenji
K
Sakamoto
Yoshiaki
Y
Katou
Shingo
S
Yoshimura
Yasunori
Y
D000328:Adult; D041321:Chromosomes, Human, X; D005188:False Negative Reactions; D005260:Female; D018628:Gene Dosage; D020440:Gene Duplication; D006801:Humans; D008297:Male; D020388:Muscular Dystrophy, Duchenne / Q000175:diagnosis* / Q000235:genetics*; D019836:Preimplantation Diagnosis / Q000379:methods*; D012107:Research Design; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods