[Diagnosing achondroplasia by single cell nested-PCR].
OBJECTIVE
To research on the reliability of diagnosing achondroplasia (ACH) on single cell level and to provide a basis for preimplantation genetic diagnosis(PGD).


METHODS
The high-frequency mutation region G380R of fibroblast growth factor receptor 3(FGFR3) gene was amplified by nested-PCR with single lymphocyte and single blastomere. The products of PCR were digested by restriction enzyme Bfm I, then the digested products were detected by 10% polyacrylamida gel electrophoresis(PAGE).


RESULTS
The amplification success rate, allele dropout rate and correct diagnosis rate of single lymphocyte's PCR were 90.4%, 8.2% and 91.8%,respectively. The amplification success rate of single blastomere was 75.4%.


CONCLUSIONS
The diagnosis of ACH by single cell nested-PCR is comparatively stable and reliable.
Zhong
Chang-gao
CG
Institute of Reproduction and Stem Cell Engineering, Reproductive and Genetic Hospital of CITIC-XIANGYA: Central South University, Changsha, Hunan, 410078 PR China.
Li
Lu-yun
LY
Lu
Chang-fu
CF
Lin
Ge
G
Fu
Jun-jiang
JJ
Luo
Ke-li
KL
Lu
Guang-xiu
GX
D000130:Achondroplasia / Q000175:diagnosis* / Q000235:genetics; D004252:DNA Mutational Analysis; D006801:Humans; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D009154:Mutation; D016133:Polymerase Chain Reaction / Q000379:methods*; D019836:Preimplantation Diagnosis; D051498:Receptor, Fibroblast Growth Factor, Type 3 / Q000235:genetics*; D012680:Sensitivity and Specificity
C496365:FGFR3 protein, human; D051498:Receptor, Fibroblast Growth Factor, Type 3