HEMOPHILIA A® GENOTYPING KIT

HEMOPHILIA A® GENOTYPING KIT is a kit developed to investigate mutations occurring in the F8 gene in DNA molecules isolated from the blood or tissue, which can perform both targeted mutation screening and conduct screening in the entire genome with next generation sequence analysis. With this kit, the “intron 22 inversion” mutation, which is the most frequent genetic change that may be responsible for the disease, can be detected in the F8 gene, while coding exons and exon-intron resultants of the F8 gene can be investigated through next generation sequence analysis method. There is a protocol required for the enzyme cut, ligation with enzymes and multiplex PCR in the part “Part A” of this kit, which is designed for the molecular diagnosis algorithm of hemophilia A disease. The “Part B” section of the kit contains materials necessary for next generation sequence analysis. This part of the kit, which was developed to enrich all targeted gene regions for next generation sequence analysis, achieves this with a single PCR condition and a PCR reaction carried out in a total of four tubes.

USAGE AREAS

HEMOPHILIA A® GENOTYPING KIT aims to identify the “intron 22 inversion” mutation in two sections in accordance with the diagnostic algorithm of the hemophilia A. It is also used to investigate the other regions of the gene rapidly, cheaply and reliably through a next generation sequence analysis method if this common mutation cannot be detected. With this kit, the Hemophilia A can be detected molecularly in 85-95% of all cases.

ORDER PRODUCT

Description

Hemophilia A is a rare hereditary hemorrhage disorder characterized by X-linked recessive inheritance that develops as deficiency of Factor VIII and is manifested by intraarticular (hemarthrosis) and intramuscular (hematoma) hemorrhages. Hemophilia A occurs one in approximately 5,000-10,000 male births. The severity of the disease is higher among the youngest patients and the clinical symptoms are experienced more severely. The disease is mostly transmitted to male children from carrier women. About one-third of cases may occur with spontaneous de-novo mutations without a family history.

The severity of hemorrhage findings is directly related to the degree of deficiency of factor VIII. Patients with factor activity (1% exhibit “severe hemophilia” clinical symptoms, while those with 1-5% exhibit “moderate hemophilia” clinical symptoms, and those with) 5% exhibit “mild hemophilia” clinical symptoms.

The molecular changes in the F8 gene, which are responsible for hemophilia A, can be divided into 3 groups:

1. Large gene rearrangements

2. Intragenic deletions or insertions

3. Single nucleotide changes

Around 5,000 variants have been reported in the Hemophilia A mutation databases. Because of the molecular pathologies in all these classes, the disease may take a severe form. However, the inversion including the F8 intron 22 has been found to be responsible for about 40-50% of severe hemophilia A cases. The most common mutation mechanism, independent of the degree of severity, may be determined as single base changes for hemophilia A.

ORDER PRODUCT

Additional information

TECHNICAL SPECIFICATIONS PART A

Catalogue Number M-HEM-01
Main Material DNA
Number of Tests 25 Reactions
Transportation conditions With dry ice and cold chain
Storage -20°C
Method Enzyme Cutting
Kit Content INV22 PCR Miz, INV22 Enzyme-I, INV22 Buffer-I, INV22 Enzyme-II, INV22 Buffer-II, INV22 Primer-I, INV22 Primer-II , INV22 Primer-III, INV22 Primer-IV

TECHNICAL SPECIFICATIONS PART B

Catalogue Number M-HEM-02
Main Material DNA
Number of Tests 25 Reactions
Transportation conditions With dry ice and cold chain
Storage -20°C
Sequence Type NGS
Kit Content Sequencing PCR Mix, Sequencing Primer Mix-I, Sequencing Primer Mix-II, Sequencing Primer Mix -III, Sequencing Primer Mix-IV