Evaluation of denaturing high‐performance liquid chromatography (DHPLC) in the screening of mutations in hemophilia B patients

Abstract

D in other cell types, e.g. keratinocytes [6] and osteoblasts [7]. Elevated PAI-1 levels have been implicated as a risk factor for cardiovascular events including myocardial infarction and stroke [8]. Patients with ESRD are known to have impaired fibrinolysis [9], in part, due to increased circulating levels of active PAI-1 [10].In summary, the expression of PAI-1, but not other gene products, was markedly reduced in paricalcitol vs. calcitrioltreated cells at concentrations of paricalcitol (4 nM) that are observed in the sera of HD patients. Because elevated PAI-1 levels are associated with increased cardiovascular mortality in patients undergoing HD, reduced expression of PAI-1 could contribute to the survival advantage observed in HD patients treated with paricalcitol. Determining PAI-1 levels in ESRD patients treated with paricalcitol and calcitriol is necessary to confirm these findings.

Molecular genetic analysis of 1053 Danish individuals with clinical signs of familial hypercholesterolemia

“…The mutational detection rate was 34.1 and 51.3% for the probands and the relatives, respectively, which is comparable to what has been found in other studies (22,(31)(32)(33)(34). Even though the mutational detection rate is high when using DHPLC with sensitivity above 95% (35), this study did not confirm mutations in all probands. This might in part be due to the inclusion criteria, as Jensen et al (16) reported a 80% mutation detection rate when using strict inclusion criteria's with a P-Chl of 10.6 mmol/l and the presence of extra-vascular lipid deposits in more than 85.6% of the patients.…”

Molecular characterization of factor IX gene mutations in 53 patients with haemophilia B in India

“…It is possible that the sensitivity for mutation detection wase xceededf or exon b+cP CR (513bp) as the optimalfragment size for mutation detection by CSGE is between 200-450bp(2). This sensitivity is comparable to thatr eported ford enaturing gradient gel electrophoresis (90.9%), single strand conformation polymorphisma nalysis (90.4%)but less than the 100% reported for denaturing high performancel iquid chromatography, at echnique thati sn ot commonlyavailable (6)(7)(8).…”

Role of Molecular Genetics in Hemophilia: From Diagnosis to Therapy

“…In contrast, the large size and complexity of the F8 gene necessitates amplifications of genomic DNA in over 30 fragments to cover the target regions. 19 Various mutation screening techniques can be used to screen PCR products of F8 or F9 genes, such as single-strand conformation polymorphism, 20 denaturing gradient gel electrophoresis, 21 conformationsensitive gel electrophoresis (CSGE), 19 and denaturing high pressure liquid chromatography 22 with sensitivities ranging from 80 to 98%. Abnormal PCR product profiles are sequenced to identify the nucleotide change.…”