A rapid sample method for HLA haplotype typization. A preliminary study on celiac patients

Area of Science:

• Immunogenetics
• Molecular Biology
• Gastroenterology

Background:

• The human leukocyte antigen (HLA) system is crucial for immune regulation and is linked to various diseases, including celiac disease.
• Genetic susceptibility to celiac disease is associated with HLA-DQB1*0201 and HLA-DQB1*0302 alleles (serologic groups -DQ2 and -DQ8).
• Early identification of these HLA genotypes is vital for diagnosing celiac disease.

Purpose of the Study:

• To determine the HLA haplotype in 64 Sardinian celiac disease patients.
• To evaluate a novel, less invasive oral mucosa brushing method for sample collection and storage.
• To assess the feasibility of this method for widespread population studies and early disease diagnosis.

Main Methods:

• Developed a less traumatic oral mucosa brushing technique for sample collection, enabling easier storage.
• Extracted DNA using Van Embden's method from collected samples.
• Utilized Polymerase Chain Reaction (PCR) with 8 specific primers for low-resolution HLA-DQB1 allele amplification.

Main Results:

• Out of 64 patients, 26 were HLA-DQB1*02 homozygotes.
• 28 patients were HLA-DQB1*02 heterozygotes.
• 10 samples were negative for the HLA-DQB1*02 allele, consistent with previous findings.

Conclusions:

• The developed oral brushing method is a reliable technique for HLA haplotype determination in non-clinical settings.
• This method facilitates early diagnosis of chronic pathologies associated with specific HLA groups.
• The technique is suitable for large-scale population studies to identify individuals with specific HLA genotypes.