Application of DHPLC for mutation detection of the fibrillin-1 in patients with Marfan syndrome / 中华检验医学杂志

ABSTRACT

Objective To detect fibrillin-1 gene (FBN1) mutations in patients with Marfan syndrome (MFS) by denaturing high-pedormance liquid chromatography (DHPLC) and DNA sequencing. Methods Genomic DNA was extracted from whole blood sample of 22 MFS patients. All 65 exens of FBN1 were amplified by polymerase chain reaction(PCR) respectively. Mutations were screened by DHPLC followed by DNA sequencing of the PCR products which showed different DHPLC profiles from the normals. Results Ten mutations of the FBN1 were found in 9 MFS patients. The mutations comprised four missense[5015G > C(C1672S),5309G > A(C1770Y),7241G > A(A2414G) and 7769G > A(C2590Y)], four nonsense [3295G > T ( E1099X ), 430"/insTCGT (G1441X), 4621C > T ( R1541X ) and 8080C > T (A2694X)], and two splice site mutations (IVS29 + 4A > T and IVSSO + 1G > A). Conclusion It is suggested that DHPLC coupled with DNA sequencing is an efficient method for the detection of FBN1 gene mutations, and it may be useful in diagnosis of MFS.