[Single cell analysis of some deletion in dystrophin gene exons and gender determination by 3-plex nested PCR].
OBJECTIVE
To set up a technique of single lymphocytes 3-plex nested PCR for dystrophin and SRY gene, and to evaluate the possibility of using this technique for preimplantation genetic diagnosis(PGD) of deleted Duchenne muscular dystrophy (DMD) with family history.


METHODS
Fifty single lymphocytes of a normal male and fifty of a normal female were obtained for detecting dystrophin gene(exon 51, exon 19, exon 48) and SRY gene by 3-plex nested PCR.


RESULTS
In the group of exon 51/exon 19/SRY, the amplification rates of exon 51, exon 19 and SRY in male were 96%, 94% and 94%; the amplification rates of exon 51 and exon19 in female were 94% and 94%, respectively. In the exon 48/exon 19/SRY group, the amplification rates of exon 48, exon 19 and SRY in male were 92%, 90% and 94%, the amplification rates of exon 48, exon 19 in female were 94% and 92%, respectively.


CONCLUSIONS
The technique of single lymphocytes 3-plex nested PCR for dystrophin and SRY gene established in this study is highly sensitive, specific and reliable, and is suitable for PGD of deleted DMD with family history.
Huang
Wen
W
Department of Neurolgy, First Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong, 510080 PR China.
Zhang
Cheng
C
Xie
You-mei
YM
Chen
Song-lin
SL
Jiao
Ze-xu
ZX
Zhou
Can-quan
CQ
Zhang
Wei-xi
WX
Lu
Xi-lin
XL
D016189:Dystrophin / Q000235:genetics*; D005091:Exons / Q000235:genetics*; D005260:Female; D006801:Humans; D008297:Male; D016133:Polymerase Chain Reaction / Q000379:methods*; D019836:Preimplantation Diagnosis / Q000379:methods; D015203:Reproducibility of Results; D017384:Sequence Deletion; D019849:Sex Determination Processes
D016189:Dystrophin