Genotyping of Hemochromatosis-Associated Mutations in the HFE Gene by PCR-RFLP and a Novel Revers Hybridization Method

Koeken, Ankie; Cobbaert, Christa; Quint, Wim; Doorn, Leen–Jan van

doi:10.1515/cclm.2002.021

Cited by 10 publications

(7 citation statements)

References 19 publications

(12 reference statements)

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“…Advanced read-out Mass spectrometry based, capillary electrophoresis, chip based n/a n/a n/a + [174][175][176][177][178][179] Reverse hybridization assay Multiplex PCR amplification followed by reverse hybridization n/a n/a n/a + [21,150,180,181] Novel extraction methods Dried blood spots, whole-blood PCR n/a n/a n/a ++ [38,158,182] Whether they are screened with the above procedure depends upon their age, health status, and the attitude of the family.…”

Section: Family Screeningmentioning

confidence: 99%

EASL clinical practice guidelines for HFE hemochromatosis

Ji-don¹

2010

Journal of Hepatology

486

167

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Iron overload in humans is associated with a variety of genetic and acquired conditions. Of these, HFE hemochromatosis (HFE-HC) is by far the most frequent and most well-defined inherited cause when considering epidemiological aspects and risks for iron-related morbidity and mortality. The majority of patients with HFE-HC are homozygotes for the C282Y polymorphism [1]. Without therapeutic intervention, there is a risk that iron overload will occur, with the potential for tissue damage and disease. While a specific genetic test now allows for the diagnosis of HFE-HC, the uncertainty in defining cases and disease burden, as well as the low phenotypic penetrance of C282Y homozygosity poses a number of clinical problems in the management of patients with HC. This Clinical Practice Guideline will therefore, focus on HFE-HC, while rarer forms of genetic iron overload recently attributed to pathogenic mutations of transferrin receptor 2, (TFR2), hepcidin (HAMP), hemojuvelin (HJV), or to a sub-type of ferroportin (FPN) mutations, on which limited and sparse clinical and epidemiologic data are available, will not be discussed. We have developed recommendations for the screening, diagnosis, and management of HFE-HC.

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Section: Family Screeningmentioning

confidence: 99%

EASL clinical practice guidelines for HFE hemochromatosis

Ji-don¹

2010

Journal of Hepatology

486

167

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“…study that compared three methods of PCR, two with mutation-specific primers and one with a novel PCRbased reverse hybridization line probe assay. This study showed a 6% underestimation by standard PCRrestriction fragment length polymorphism (RFLP) methods [7]. Future studies should consider the use of the most accurate methods for HFE polymorphism screening.…”

Section: Discussionmentioning

confidence: 75%

“…This is an indirect method for diagnosis and the method originally used to determine gene frequencies. Recently, by use of a direct method of DNA analysis, three specific mutations in the HFE gene associated with iron overload have been identified [7]. These include one ''major'' hemochromatosis gene mutation, known as C282Y, and two ''minor'' mutations at H63D and S65C [8].…”

Section: Introductionmentioning

confidence: 99%

Identification of hemochromatosis gene polymorphisms in chronically transfused patients with sickle cell disease

Jeng¹,

Adams-Graves²,

Howard³

et al. 2003

American J Hematol

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Three polymorphic gene mutations in the human hemochromatosis (HFE) gene (C282Y, H63D, S65C) are associated with non-transfusion-related iron overload in Caucasians. More recently, these mutations have also been identified in African-Americans. However, the prevalence of HFE gene mutations in African-Americans with sickle cell disease (SCD) has not been described. The presence of these mutations in this population is particularly important, because patients with SCD may be placed on chronic red cell transfusion therapy and are thus at further risk for iron overload. Thus, we attempted to establish the gene mutation prevalence in African-Americans with SCD, to compare this frequency with published gene frequencies in African-Americans, and to evaluate their significance with regard to transfusion-related iron overload. Eighty-nine African-American patients with SCD, all of whom were receiving chronic red cell transfusion therapy, were screened by DNA analysis for the three HFE gene mutations. Two patients were heterozygous for the C282Y HFE mutation (2.3%), six were heterozygous for the H63D mutation (6.8%), none carried the S65C mutation (0.0%), and no homozygous or compound heterozygous subjects were identified. The prevalence of C282Y and H63D in the SCD population was similar to that observed in the general African-American population. In addition, there was no increased mutation prevalence when comparing those SCD patients on chronic transfusion therapy who had ferritin levels greater than 2,500 ng/mL and those less than 2,500 ng/mL. This study represents the first identification of the known HFE gene mutations by DNA analysis in the SCD population. We conclude that the presence of recognized HFE coding region mutations do not seem to have an impact on the degree of iron overload in patients with SCD receiving chronic transfusion therapy.

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“…The PCR-RFLP method is a widely used method for genotyping of gene (Hao et al, 2007;Jauk et al, 2003;Koeken et al, 2002;Sironi et al, 2010;Wang et al, 2010).…”

Section: Discussionmentioning

confidence: 99%

Genotyping of flavin-containing mono-oxygenase 3 (FMO3) gene by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and mismatch amplification mutation assay (MAMA-PCR) in chickens

Hamid¹,

Wang²,

Zhao

2012

Afr. J. Biotechnol.

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Fishy smelling off-flavour of chicken meat is caused by a mutation (FMO3 c.985A>T) in exon 7 of flavincontaining mono-oxygenase 3 (FMO3) gene. Comparative analysis of the causative mutation in FMO3 gene in different chickens using two different methods was reported. Polymerase chain reactionrestriction fragment length polymorphism (PCR-RFLP) method was based on general primers, whereas another employed method was the mismatch amplification mutation assay (MAMA) primers. Seventy (22.88%) of the 306 samples were genotyped using PCR-RFLP, 194 (63.40%) of the 306 samples were genotyped using MAMA-PCR and 42 (13.72%) of the 306 samples were genotyped by both of PCR-RFLP and MAMA-PCR and genotyping data were validated by DNA sequencing. The results show that the genotyping accuracy for PCR-RFLP was 90.48% in AA, 85.42% in AT and 95.35% in TT and 100% in AA, 98% in AT and 96% in TT for MAMA-PCR. This study suggests that the MAMA-PCR is more effective, accurate, easy, fast, efficient and reliable than PCR-RFLP.

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Genotyping of Hemochromatosis-Associated Mutations in the HFE Gene by PCR-RFLP and a Novel Revers Hybridization Method

Cited by 10 publications

References 19 publications

EASL clinical practice guidelines for HFE hemochromatosis

EASL clinical practice guidelines for HFE hemochromatosis

Identification of hemochromatosis gene polymorphisms in chronically transfused patients with sickle cell disease

Genotyping of flavin-containing mono-oxygenase 3 (FMO3) gene by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and mismatch amplification mutation assay (MAMA-PCR) in chickens

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