F8 SegCheck Kit v1.2

Features and Advantages:

• • Disease gene tracing in the pedigree
  • Clearing the ambiguity of the embryonic sample contamination with the maternal sample
  • Clearing the ambiguity of the sample contamination with another sample
  • Confirming the authenticity of the embryonic sample and identifying the sample swapping error
  • Determining the sample’s gender
  • Identifying the sample
  • Primary chromosomal aneuploidy screening
  • Diagnosing and distinguishing embryonic samples in twins
  • Diagnosing uniparental disomy (heterodisomy or isodisomy)
  • Diagnosing the crossing-over phenomenon in HLA examination and disease creation
  • Diagnosing embryonic mosaicism
  • Gonadal mosaicism (more in X-linked disorders)
  • Confirming mutation diagnosis results using direct methods like DNA Sequencing, ARMS, MLPA, Gap PCR, and NGS

The Importance of Using ™SegCheck Kits: In most countries, prenatal testing for chromosomal aneuploidies is performed routinely. Most common methods (i.e., first and second trimester ultrasound and biochemical screening) have limitations due to the presence of false positives or false negatives in diagnosing chromosomal aneuploidies. QF-PCR has long become a preferred method for the rapid diagnosis of common chromosomal aneuploidies. However, techniques like QF-PCR and karyotype must be requested by specialists and performed on selected embryonic samples. Aware of these shortcomings, Kowsar Biotechnology has produced the ™SegCheck kits designed for more than 60 diseases allowing laboratories to determine any disease along with primary aneuploidy screening using QF-PCR method without the need for sampling and additional costs. While NIPT (Non-Invasive Prenatal Testing) tests solely examine aneuploidy for the tested embryonic sample, the ™SegCheck kit can evaluate both the disease gene and aneuploidies for any embryonic sample undergoing prenatal diagnostic testing. Indeed, each ™SegCheck kit contains specific STR markers from chromosomes 13, 18, 21, and sex chromosomes along with disease-specific STR markers.

Selected STR markers are 4 nucleotide repeats chosen to reduce analysis and simplify interpretation. Alleles from these STR markers can assist in identifying the sample, ruling out maternal cell contamination, and many other benefits in a single test. Thus, these chromosomal markers can also be used to examine the presence of aneuploidy for the tested chromosomes. If aneuploidy is suspected, the sample can be analyzed with more markers using the QF-PCR method, such as the AneuQuick™ Kit v3.2 or AneuQuick™ Extra Plus Kit v1.2.

Every genetic diagnostic laboratory performing prenatal diagnostics faces issues like sample mixing and maternal cell contamination. These errors can lead to incorrect diagnosis and false-negative screenings. ™SegCheck can assist any medical genetic laboratory conducting prenatal diagnostics. As these kits have markers inside or beside the specific disease gene and are segregated, these markers can be used for segregation tracking and linkage analysis. Drawing haplotypes confirms the inheritance of disease gene mutations in embryos.

For example, suppose parents have a similar mutation, and due to allelic dropout or ADO during direct mutation diagnosis in the embryo, it appears homozygous, indicating the mutation in haplotype drawing and its analysis in a family shows it is actually heterozygous due to ADO, where the other gene is not amplified. Consequently, the embryo perceived as a carrier would have been discarded. The reason ™SegCheck kits can detect ADO is that each kit contains between 4 to 8 STR markers, and it’s improbable for a sample to exhibit ADO for all these markers.

This methodology used in these kits will be valuable for most molecular medical genetics laboratories conducting prenatal diagnosis. However, they could also be applied by other researchers for different purposes like gene segregation, inheritance, or even mosaicism.

Hemophilia A, the most common inherited bleeding disorder linked to sex with a prevalence of 1 in 5000 to 6000 male births, results from mutations in the Factor 8 (F8) gene causing a defect in Factor VIII coagulation protein. The disease is predominantly seen in males and exists in mild, moderate, and severe forms. Newly reported cases are mostly due to new mutations. This gene is located on the long arm of chromosome X (Xq28).

The F8 SegCheck™ Kit v1.2 is a molecular diagnostic kit that quickly diagnoses carriers and those affected by Factor VIII coagulation deficiency, commonly known as Hemophilia A. Primarily based on STR markers and QF-PCR technique, the kit employs segregation analysis and haplotyping to determine whether the embryonic sample is affected or a carrier. Using STR markers along with direct mutation detection, prenatal diagnosis becomes more accurate and reliable. Along with detecting Factor VIII coagulation defect, the F8 SegCheck™ v1.2 kit includes autosomal STR markers for primary screening of common chromosomal disorders (3 markers for chromosome 21, 2 markers for chromosome 18, and 2 markers for chromosome 13) and sex determination.

The diagnostic F8 SegCheck™ v1.2 kit is based on 16 markers which include 5 high heterozygosity STR markers upstream and downstream of the F8 gene and 11 markers for primary screening of common chromosomal disorders and sex determination.

This kit is compatible with operating on DNA samples extracted from blood, amniotic fluid (AF), chorionic villus sampling (CVS), and DBC cards (DNA storage cards from Kowsar Biotech), blood lysis buffer BLB, amniotic cell lysis buffer AFL, and CVL lysis buffer for chorionic villi. For more information on product usage and analysis of results, refer to the manual.

DXF8SU6.1، DXF8SU6، DXF8SD1.6، DXF8D8.4، DXF8SD15.9، D21S1809، D21S1414، D21S1442، D21S1411، D18S391، D18S1002، D13S252، DXS7132، D13S634، SRY و AMXY.

The F8 SegCheck™ v1.2 kit is compatible with 5-color capillary electrophoresis systems like the ABI PRISM 3130/3130xl/3500/3500xL Genetic Analyzers with 30, 50, or 80 cm capillary sizes.