Celiac Disease (Coeliakie) PCR Kit

The Celiac Disease Pcr Kit: CeliaSCANCeliac Disease PCR

Goffin Molecular Technologies introduced the solution to  CeliaSCAN kit (developed by TubaScan Ltd., NL), an easy to use multiplex real-time PCR Assay and Melting Curve Analysis for the in vitro detection of strongly Celiac Disease-associated human leukocyte antigen (HLA)-DQ2.5, HLA-DQ2.2, and HLA-DQ8.  Celiac Disease PCR

Celiac Disease Celiac Disease PCR

Celiac disease is an autoimmune disease presenting with or without symptoms such as chronic diarrhea, nutritional malabsorption, weight loss, unexplained iron deficiency anemia, osteoporosis or neurological symptoms. Celiac disease is caused by an inappropriate immune response to dietary gluten, found in wheat, rye, and barley.

Often, in the small bowel villous atrophy is present and intraepithelial lymphocyte (IEL) levels are increased.

The worldwide prevalence is ~1%, most celiac disease is undetected.

90% of patients with celiac disease express the HLA-DQA1*05, HLA-DQB1*02 heterodimer (HLA-DQ2.5). Of the remaining 10% most carry the HLA-DQA1*03, HLA-DQB1*03:02 heterodimer (HLA-DQ8). Some of the patients with celiac disease express the HLA-DQA1*02, HLA-DQB1*02 heterodimer (HLA-DQ2.2).

Absence of these haplotypes is a strong negative predictor of celiac disease.

Individuals homozygous for HLA-DQ2.5 have the highest genetic risk for celiac disease

CeliaScan Test Principle:Celiac Disease PCR

Extraction of DNA from buccal swabs or whole blood is not included.

  • Multiplex real-time PCR and Melting Curve assay
  • Simultaneous amplification of allele specific parts in exon 2 of both the HLA-DQA1 and HLA-DQB1 genes
  • 6 primer pairs detect 6 alleles in 3 reactions
  • Two additional primer pairs detect monomorphic parts of the HLA-DRA and HLA-F genes and serve as amplification controls
  • SYBR Green fluorophore binds to double-stranded DNA and emits green light if bound
  • PCR reaction followed by melting curve analysis
  • Each peak generated represents the characteristic melting temperature (Tm) of an amplicon where the DNA is 50% double-stranded and 50% single-stranded

Analysis and Reporting of Results:

  • Highly advanced online software tool (ClinicaGeno, Ltd., UK)
  • Experiment files uploaded to server
  • Analysis of results by software algorithm, all peaks are detected and translated into the correct genotype
  • The report denotes all HLA-DQA1 and HLA-DQB1 at-risk alleles, the complete at risk genotype and zygosity status.

Benefits:

  • Fast, 1.5 hrs Assay including Analysis & Reports
  • In-house comparison with Single-Strand Conformation Polymorphism method: Sensitivity ~100% and specificity~100%
  • Able to distinguish HLA-DQ2.2 from HLA-DQ2.5
  • Able to distinguish HLA-DQ2.5 homozygotes from HLA-DQ2.5 heterozygotes
  • Fully automated online results analysis tool, therefore no human interpretation of data is needed

Click here to see the comparison of the CeliaScan to an in-house SSCP/HD method