Diagnosing achondroplasia by single cell nested-PCR.
- Author:
Chang-gao ZHONG
1
;
Lu-yun LI
;
Chang-fu LU
;
Ge LIN
;
Jun-jiang FU
;
Ke-li LUO
;
Guang-xiu LU
Author Information
- Publication Type:Journal Article
- MeSH: Achondroplasia; diagnosis; genetics; DNA Mutational Analysis; Humans; Molecular Diagnostic Techniques; methods; Mutation; Polymerase Chain Reaction; methods; Preimplantation Diagnosis; Receptor, Fibroblast Growth Factor, Type 3; genetics; Sensitivity and Specificity
- From: Chinese Journal of Medical Genetics 2003;20(3):228-231
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo research on the reliability of diagnosing achondroplasia (ACH) on single cell level and to provide a basis for preimplantation genetic diagnosis(PGD).
METHODSThe 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).
RESULTSThe 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%.
CONCLUSIONThe diagnosis of ACH by single cell nested-PCR is comparatively stable and reliable.
