Evaluation of DHPLC in the analysis of hemophilia A

Oldenburg, Johannes; Ivaškevičius, Vytautas; Rost, S.; Fregin, Andreas; White, Karen L.; Holinski‐Feder, Elke; Müller, C. R.; Weber, Bernhard H. F.

doi:10.1016/s0165-022x(00)00150-0

Cited by 76 publications

(68 citation statements)

References 19 publications

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“…These data are in agreement with the results of earlier investigations in the Italian population and with other studies in a sizeable series of patients. 9,12,25 We identified 28 novel ABCR mutations, confirming and further highlighting the high heterogeneity of the ABCR gene in STGD. The frequent report of novel mutations is probably related to the enormous number of exons in the ABCR gene, which favours rearrangement in the DNA sequence.…”

Section: Discussionsupporting

confidence: 62%

Novel mutations in of the ABCR gene in italian patients with Stargardt disease

Passerini

Sodi

Giambene

et al. 2009

Eye

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Purpose: Stargardt disease (STGD) is the most prevalent juvenile macular dystrophy, and it has been associated with mutations in the ABCR gene, encoding a photoreceptor-specific transport protein. In this study, we determined the mutation spectrum in the ABCR gene in a group of Italian STGD patients. Methods: The DNA samples of 71 Italian patients (from 62 independent pedigrees), affected with autosomal recessive STGD, were analysed for mutations in all 50 exons of the ABCR gene by the DHPLC approach (with optimization of the DHPLC conditions for mutation analysis) and direct sequencing techniques. Results: In our group of STGD patients, 71 mutations were identified in 68 patients with a detection rate of 95.7%. Forty-three mutations had been already reported in the literature, whereas 28 mutations had not been previously described and were not detected in 150 unaffected control individuals of Italian origin. Missense mutations represented the most frequent finding (59.2%); G1961E was the most common mutation and it was associated with phenotypes in various degrees of severity. Conclusions: Some novel mutations in the ABCR gene were reported in a group of Italian STGD patients confirming the extensive allelic heterogeneity of this geneFprobably related to the vast number of exons that favours rearrangements in the DNA sequence.

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Section: Discussionsupporting

confidence: 62%

Novel mutations in of the ABCR gene in italian patients with Stargardt disease

Passerini

Sodi

Giambene

et al. 2009

Eye

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“…HA affects 1 in 5000 males [1][2][3] . Generally males are affected and females are carriers of the disease.…”

Section: Introductionmentioning

confidence: 99%

“…The severity of the disease depends on the level of normal F8 activity and is classified as severe cases (<1%), moderate (1-5%), and mild (>5-30%). Among all the HA patients, about 50% have severe form of the disease, while 10% are moderate and 40% are mild [1][2][3][4] . The type of bleeding symptoms is related to the severity of the disease.…”

Section: Introductionmentioning

confidence: 99%

“…Patients with severe haemophilia A have spontaneous bleeding, but patients with moderate disorder experience infrequent spontaneous bleeding and their bleeding is typically after trauma or surgery. Mild patients have bleeding after surgery or trauma 3,4,6,7 . Anti-FVIII antibodies are the most serious complication that arises in response to infusion of factor VIII concentrates 3,4 .…”

Section: Introductionmentioning

confidence: 99%

“…Mild patients have bleeding after surgery or trauma 3,4,6,7 . Anti-FVIII antibodies are the most serious complication that arises in response to infusion of factor VIII concentrates 3,4 . The frequency of inhibitor development in severe patients (20-40%) is higher than mild or moderate (1-13%) HA cases.…”

Section: Introductionmentioning

confidence: 99%

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Evaluation of intron 22 and intron 1 inversions of the factor 8 gene using an inverse shifting PCR method in severe haemophilia A patients

Roozafzay¹,

Kokabee²,

Zeinali³

et al. 2013

ScienceAsia

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Haemophilia A (HA) is an X-linked recessive bleeding disorder caused by several mutations in the F8 gene, whose inversion causes most of the mutation in 40% of severe HA patients. Methods such as long distance PCR and southern blot have been exploited to analyse intron 22 inversion in the F8 gene. For this purpose, we used an inverse shifting PCR assay, a novel genetic diagnostic method, for the detection of intron 22 inversion in severe HA patients. A screen for the presence of intron 22 and intron 1 inversions at the F8 gene in 30 Iranian severe HA patients revealed that 47% of patients showed intron 22 inversion (40% type 1 and 7% type 2), while 7% of patients had intron 1 inversion. Among the patients carrying the inversion, 12% developed inhibitors. Inverse shifting PCR is a precise method for assessment of rearrangements related to int 22h and int 1h in patients and carriers of haemophilia A. Since the previous methods of assessing inversion are time-consuming and demanding, this method can be a good replacement.

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Small FVIII gene rearrangements in 18 hemophilia A patients: Five novel mutations

et al. 2005

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Hemophilia A (HA) is a disorder caused by mutations of the FVIII gene, which is located on the tip of the long arm of the X chromosome. In a cohort of 18 unrelated Italian patients affected with HA of varying severity, we performed mutational screening of the gene by denaturing high-performance liquid chromatography (DHPLC) and direct sequencing of abnormal peaks. We identified five novel mutations and 9 previously reported DNA alterations. Two of the 9 previously reported alterations were each common to 3 unrelated patients. Six different mutations were characterized as missense alterations, while 8 were non-missense mutations. Among the new gene alterations, one created a stop codon, one consisted of an out-of frame deletion, and one was a splice-site mutation. The last two were missense alterations. In an attempt to better understand the causative effect of the mutations and the clinical variability of the patients, we investigated the consequences of each missense mutation and visualized the effect of the amino acid change on structural FVIII models. Am.

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Evaluation of DHPLC in the analysis of hemophilia A

Cited by 76 publications

References 19 publications

Novel mutations in of the ABCR gene in italian patients with Stargardt disease

Novel mutations in of the ABCR gene in italian patients with Stargardt disease

Evaluation of intron 22 and intron 1 inversions of the factor 8 gene using an inverse shifting PCR method in severe haemophilia A patients

Small FVIII gene rearrangements in 18 hemophilia A patients: Five novel mutations

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