Molecular detection of Toll-like receptors 1, 4, and 5 genes among patients with appendicitis
Abstract
Background & Objective: Acute appendicitis is the predominant etiology of emergency cases on a global scale, and it may have serious consequences. If there is no precise diagnosis. Toll-like receptors (TLRs) are a crucial group of receptors that form the initial barrier against infections. The study aimed to investigate the molecular structure of Toll-like receptor (TLRs) genes by conventional Polymerase Chain Reaction) PCR) and deoxyribonucleic acid (DNA) sequencing in acute appendicitis patients and investigate whether any alteration in this gene may act as a risk factor for appendicitis.
Methodology: A case-control investigation was conducted at Al-Sadr Teaching Hospital and Al-Shifaa Teaching Hospital in Basrah, Iraq, from July to November 2023. The study involved 20 blood samples from 15 patients previously diagnosed with appendicitis; and 5 healthy individuals included as a control group. We randomly selected six samples from patients with appendicitis, with two samples for each primer, for DNA sequencing.
Results: The results of conventional PCR and electrophoresis showed different molecular weights at different temperatures for TLR1, TLR4, and TLR5. DNA sequencing results showed several different mutations for each TLRs gene primer for both reverse and forward when compared by the Basic Local Alignment Search Tool (BLAST) to sequences from the National Centre for Biotechnology Information (NCBI). Six new mutations were recorded in the gene bank (NCBI), two mutations per TLR. The study found that TLR1’s forward primer had two mutations, while the reverse primer had four. Four alterations were found in TLR4, while one in TLR5 was found in the reverse primer.
Conclusion: Through altering the immune system’s reaction to pathogenic stimuli, excessive immunological activation, and inflammation, genetic differences in TLRs may influence the pathophysiology or outcome of disease. These factors can also contribute to the development of appendicitis.
Abbreviations: AA, Acute Appendicitis; AHC, apparently healthy controls, “BLAST, Basic Local Alignment Search Tool”; CD, cluster of differentiation; EDTA, ethylene diamine tetra acetic acid; GBD, Global Burden of Disease Study; GC, gastric cancer; GIT, gastrointestinal tract; IBD, inflammatory bowel disease; LPS, lipopolysaccharide; “MyD88, myeloid differentiation primary response 88”; NCBI, National Center for Biotechnology Information; PRRs, “Pathogen Recognition Receptors; PAMPs Pathogen Associated Molecular patterns”; SNPs, single-nucleotide polymorphisms; TE, Tris-EDTA; TIR, Toll-interleukin (IL)-1 receptor.
Keywords: Appendicitis, TLR1, TLR4, TLR5, PCR, DNA sequencing, Molecular Detection
Citation: Shaker ZN, Mahdi DS, AlSaimary IE. Molecular detection of Toll-like receptors 1, 4, and 5 genes among patients with appendicitis. Anaesth. pain intensive care 2024;28(6):1050-1060; DOI: 10.35975/apic.v28i6.2622
Received: August 22, 2024; Reviewed: October 06, 2024; Accepted: October 18, 2024
