Multiplex ligation‐dependent probe amplification as first mutation screening for large deletions and duplications in haemophilia

Belvini, Donata; Salviato, Roberta; Radossi, Paolo; Tagariello, Giuseppe

doi:10.1111/hae.13143

Cited by 7 publications

(4 citation statements)

References 28 publications

(32 reference statements)

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“…This technique quantifies the copy number of targeted genes in a patient's DNA compared to a control's and has proved to be effective in detecting deletions and duplications in suspected HA female carriers when traditional screening had failed. 5 MLPA revealed a large F8 deletion encompassing exons 1-9 in the propositus ( Figure 2B) and her mother, confirming that they were carriers of SHA. Large deletions occur in 3% of SHA cases 2 and a similar large deletion encompassing exons 1-9 has previously been reported in a subject with SHA and FVIII inhibitor.…”

Section: The Diagnosis Of a Haemophilia A Carrier Over 2 Decadesmentioning

confidence: 65%

The diagnosis of a haemophilia A carrier over 2 decades

et al. 2020

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Section: The Diagnosis Of a Haemophilia A Carrier Over 2 Decadesmentioning

confidence: 65%

The diagnosis of a haemophilia A carrier over 2 decades

et al. 2020

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“…The mRNA length of F9 gene is 2804 bp containing 8 exons and encoding 461 amino acids. At present, Multiplex ligation dependent probe amplification (MLPA), Sanger sequencing and high throughput sequencing techniques have been used in gene test for HB [9–11] and they have their own advantages. MLPA can detect large deletion or duplication mutations in F9 gene, but cannot find point mutations with high mutation rate.…”

Section: Discussionmentioning

confidence: 99%

Genetic analysis of a hemophilia B family with a novel F9 gene mutation

Li³

et al. 2019

Medicine

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At present, there are no effective methods for the treatment of hemophilia B, and it has high mortality and disability. Therefore, it is very important for the carriers to carry out genetic counseling and make prenatal diagnosis. In this study, we made gene and prenatal diagnoses in a family with a novel F9 gene mutation, and report a novel F9 gene mutation. All exon sequences and flanking sequences of F9 gene were analyzed by Sanger sequencing in the proband; and then according to the F9 gene mutation in the proband, the F9 gene sequencing was performed on the family members. Based on the above results, the pathogenic mutation in F9 gene was finally identified, which was used for prenatal diagnosis. Sanger sequencing revealed c.1232G>C [p.Ser411Thr] mutation in F9 gene in the proband. c.1232G>C heterozygous mutation was also found in the proband's mother and grandmother, but male family members without hemophilia B had no this mutation. The analyses of amniotic fluid samples indicated positive sex-determining region on Y chromosome ( SRY ), and no c.1232G>C [p.Ser411Thr] mutation in F9 gene. We identified a pathogenic mutation in F9 gene in the family, made a prenatal diagnosis for the female carrier and reported a novel F9 gene mutation.

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“…Analysis of the obtained data was performed through Chromas sequence viewer software. Finally, if no mutation was found, the gene of FIX was quanti ed by multiplex ligation-dependent ampli cation (MLPA) (MRC-Holland, Amsterdam, the Netherlands) [13].…”

Section: Sanger Sequencing Methodsmentioning

confidence: 99%

Genotype-phenotype analyses of Iranian patients with and without hemophilia B Leyden: A single-center study

Moghadam,

Manafzadeh,

Dajliry

et al. 2024

Preprint

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Background There is a high prevalence of inherited bleeding disorders in Iran, such as hemophilia A (HA) and hemophilia B (HB). This study aimed to analyze the molecular and clinical profiles of patients with HB.Methods A single-center study was conducted among patients with severe HB between March 20, 2000, and June 31, 2023. The polymerase chain reaction (PCR) amplification was used for all of the major regions, such as the promoter, the exons, the adjacent intronic regions, and the untranslated regions of the F9 gene. Finally, Sanger sequencing was performed on the PCR products.Results A total of 111 HB patients (17 with HB Leyden and 94 without HB Leyden) were enrolled in this study. The median age of the patients at the time of diagnosis was 12 months (IQR: 6 months to 60 months). A family history of hemophilia was reported in 64 (57.7%) of patients. The most common bleeding manifestations were hemarthrosis, bruising, and oral cavity bleeding. Among 94 patients without HB Leyden, 59 (62.8%) had missense, 21 (22.3%) had nonsense, and 8 (8.5%) had frameshift mutations. Moreover, the most frequent mutation in HB Leyden was c.-17 A > G in this study. Finally, two novel mutations (c. -14 T > C and c. -56 T > A) were identified in the promotor region.Conclusion The results of this study confirm that HB is caused by a wide range of molecular defects in Iran. Thus, by knowing the genotypes and phenotypes, we would be able to stratify the patients which is important in terms of their management and outcome.

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Multiplex ligation‐dependent probe amplification as first mutation screening for large deletions and duplications in haemophilia

Abstract: Owing to its simplicity, MLPA seems useful at the beginning of the molecular investigation, saving all the following steps, where positive. Single exon deletion diagnosis requires caution due to the risk of misdiagnosis, but benefits of MLPA appear to overcome the pitfalls.

Cited by 7 publications

References 28 publications

The diagnosis of a haemophilia A carrier over 2 decades

The diagnosis of a haemophilia A carrier over 2 decades

Genetic analysis of a hemophilia B family with a novel F9 gene mutation

Genotype-phenotype analyses of Iranian patients with and without hemophilia B Leyden: A single-center study

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