Genotyping of intron 22-related rearrangements of F8 by inverse-shifting PCR in Egyptian hemophilia A patients

Abou-Elew, Heba Hassan; Ahmed, Hanan H.; Raslan, Hanan; Abdel‐Wahab, M. F.; Hammoud, Rania; Mokhtar, Doha A.; Arnaout, Hanaa H.

doi:10.1007/s00277-010-1115-x

Cited by 22 publications

(24 citation statements)

References 29 publications

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“…The same tendency was found for the intron 1 inversion, which is reported with a frequency of 5% [10]. When we compare our findings with previous data from other Arab countries the frequencies of these 2 inversions are lower in our patients [11-14]. This result shows that our patients have a specific genetic profile which is confirmed by other molecular studies concerning our country [15].…”

Section: Discussionsupporting

confidence: 91%

First report of molecular diagnosis of Tunisian hemophiliacs A: Identification of 8 novel causative mutations

Elmahmoudi¹,

Khodjet‐El‐Khil²,

Wigren

et al. 2012

Diagn Pathol

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IntroductionHemophilia A is an X linked recessive hemorrhagic disorder caused by mutations in the F8 gene that lead to qualitative and/or quantitative deficiencies of coagulation factor VIII (FVIII). Molecular diagnosis of hemophilia A is challenging because of the high number of different causative mutations that are distributed throughout the large F8 gene. Molecular studies of these mutations are essential in order to reinforce our understanding of their pathogenic effect responsible for the disorder.AimIn this study we have performed molecular analysis of 28 Tunisian hemophilia A patients and analyzed the F8 mutation spectrum.MethodsWe screened the presence of intron 22 and intron 1 inversion in severe hemophilia A patients by southern blotting and polymerase chain reaction (PCR). Detection of point mutations was performed by dHPLC/sequencing of the coding F8 gene region. We predict the potential functional consequences of novel missense mutations with bioinformatics approaches and mapping of their spatial positions on the available FVIII 3D structure.ResultsWe identified 23 different mutations in 28 Tunisian hemophilia A patients belonging to 22 unrelated families. The identified mutations included 5 intron 22 inversions, 7 insertions, 4 deletions and 7 substitutions. In total 18 point mutations were identified, of which 9 are located in exon 14, the most mutated exonic sequence in the F8 gene. Among the 23 mutations, 8 are novel and not deposited in the HAMSTeRS database nor described in recently published articles.ConclusionThe mutation spectrum of Tunisian hemophilia A patients is heterogeneous with the presence of some characteristic features.Virtual slidesThe virtual slide(s) for this article can be found here:http://www.diagnosticpathology.diagnomx.eu/vs/1693269827490715

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Section: Discussionsupporting

confidence: 91%

First report of molecular diagnosis of Tunisian hemophiliacs A: Identification of 8 novel causative mutations

Elmahmoudi¹,

Khodjet‐El‐Khil²,

Wigren

et al. 2012

Diagn Pathol

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“…Our findings confirm those of previous studies. The frequency of Inv 22 in our study was 44 %, which is similar to that in populations studied in the United Kingdom [13], Spain [17], Italy [18], Argentina [19,20], India [21], Egypt [22], Iran [23], Mexico [24], Hungary [15], and Costa Rica [25]. On the basis of these results, we concluded that the incidence in different populations is similar.…”

Section: Discussionsupporting

confidence: 74%

Genotyping of Intron Inversions and Point Mutations in Exon 14 of the FVIII Gene in Iranian Azeri Turkish Families with Hemophilia A

Khaniani

Ebrahimi

Daraei

et al. 2016

Indian J Hematol Blood Transfus

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Hemophilia A (HA) is an inherited X-linked bleeding disorder caused by a variety of mutations that are distributed throughout the large FVIII gene (F8). The most common mutations in studied populations with severe HA are introns 22 and 1 inversions, gross exon deletions and point mutations in exon 14. The aim of this study was to define the frequency of these common mutations in Iranian population of Azeri Turkish in North West of Iran. Fifty patients with severe HA and forty-three female potential carriers were genotyped by inverse shifting polymerase chain reaction (IS-PCR), long-range PCR, multiplex PCR, and sequencing methods for the detection of Intron 22 and 1 inversions, gross exon deletions, and exon 14 point mutations, respectively. F8 intron 22 inversion was detected in 22 (44 %) out of 50 patients. Moreover, we detected one intron 1 inversion (2 %), and one point mutation in exon 14 (2 %). In this population, 52 % of the patients with hemophilia A did not show to carry a mutation in the analyzed regions by three mentioned methods. F8 intron 22 inversion was the major causative mutation in nearly 50 % of severe HA cases in an Azerbaijani Turkish population, which is similar to the incidence of other populations. IS-PCR is a robust, rapid, efficient, and costeffective method for the genetic analysis of patients with severe HA and for HA carrier detection, especially in developing countries.

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“…IS-PCR avoids PCR amplification through long and GC rich sequences of F8 gene duplicons associated with intron 22h and intron 1h that these sequences may be amplified in LD-PCR 18,21 . Recently, IS-PCR method has been developed 18 and applied for the detection of int 22 inversion in different studies 25,26 . In this study we used IS-PCR assay to evaluate the presence of int 22 and int 1 inversion in severe HA patients from a single centre in Isfahan, Iran.…”

Section: Discussionmentioning

confidence: 99%

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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Genotyping of intron 22-related rearrangements of F8 by inverse-shifting PCR in Egyptian hemophilia A patients

Cited by 22 publications

References 29 publications

First report of molecular diagnosis of Tunisian hemophiliacs A: Identification of 8 novel causative mutations

First report of molecular diagnosis of Tunisian hemophiliacs A: Identification of 8 novel causative mutations

Genotyping of Intron Inversions and Point Mutations in Exon 14 of the FVIII Gene in Iranian Azeri Turkish Families with Hemophilia A

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

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