Preimplantation and prenatal genetic diagnosis of aromatic L-amino acid decarboxylase deficiency with an amplification refractory mutation system-quantitative polymerase chain reaction.
OBJECTIVE
To develop a diagnostic platform for preimplantation genetic diagnosis (PGD) and prenatal genetic diagnosis (PND) to prevent births of aromatic L-amino acid decarboxylase deficiency (AADC) patients.


METHODS
Five Taiwanese families carrying AADC were enrolled. A novel technique, amplification refractory mutation system-quantitative polymerase chain reaction (ARMS-qPCR), was developed for both of PGD and PND. For PGD, blastomere biopsies of day-3 cleavage-stage embryos were subjected to ARMS-qPCR. Villi, cultured amniocytes, or both were used to confirm the PGD result; this approach could also be used as the sole method for PND after in vivo conception).


RESULTS
Unaffected live births were achieved in four of the five families, except one with ongoing PGD. The ARMS-qPCR correctly classified blastomeres (from day-3 cleavage-stage embryos) as affected (homozygous mutant), carrier (heterozygous for mutant and wild-type alleles), or normal (homozygous wild-type) within 1 working day.


CONCLUSIONS
To our knowledge, this is the first report of successful PGD of AADC. The molecular technique we devised (ARMS-qPCR) was applicable for PGD as well as PND of AADC. Furthermore, it has great potential for similar applications in other monogenic disorders.
Department of Genomic Medicine, Changhua Christian Hospital, Changhua, Taiwan.
Shou-Jen
SJ
Kuo
Gwo-Chin
GC
Ma
Shun-Ping
SP
Chang
Hsin-Hung
HH
Wu
Chih-Ping
CP
Chen
Tung-Ming
TM
Chang
Wen-Hsiang
WH
Lin
Sheng-Hai
SH
Wu
Mei-Hui
MH
Lee
Wuh-Liang
WL
Hwu
Ming
M
Chen
D000592:Amino Acid Metabolism, Inborn Errors / Q000175:diagnosis* / Q000235:genetics; D001142:Aromatic-L-Amino-Acid Decarboxylases / Q000172:deficiency / Q000235:genetics; D001757:Blastomeres; D004296:Dopa Decarboxylase / Q000235:genetics*; D005260:Female; D006580:Genetic Carrier Screening; D005820:Genetic Testing / Q000379:methods*; D006579:Heterozygote; D006720:Homozygote; D006801:Humans; D009154:Mutation; D016133:Polymerase Chain Reaction / Q000379:methods*; D011247:Pregnancy; D011296:Prenatal Diagnosis / Q000379:methods*; D013624:Taiwan
D004296:Dopa Decarboxylase; D001142:Aromatic-L-Amino-Acid Decarboxylases