Molecular sequencing and phenotyping study of chemokines CCL2, CCL5, and CXCL10 in patients with neuroinflammation
Ahmed Salim1, Ihsan AlSaimary2, Amal Adil Kasid Alsudany3
1Department of Medicine, College of Medicine, University of Basrah, Basrah, Iraq.
Introduction:
A molecular sequencing and phenotyping study was conducted on chemokines CCL2, CCL5, and CXCL10 to assess the relation to multiple sclerosis (MS).
Material And Methods:
The molecular detection study included extractions of DNA from the blood of cases and a control group, as well as amplification and confirmation of all extracted DNA. The 490-bp primer-size PCR product corresponded to CCL2, the 481-bp primer-size PCR product corresponded to CCL5, and the 310-bp primer-size PCR product corresponded to CXCL10.
Results:
DNA sequencing of CCL2, CCL5, and CXCL10 showed that there was convergence between the obtained results and data from the GenBank database (NCBI) with 99% identity (439/445) in the forward CCL2 sequence, which had a single transition mutation, GGT to GGG. In addition, the reverse CCL2 showed 99% identity (426/427) when compared with the GenBank database (NCBI), which found a single deletion mutation, TGA to GGG. There was convergence between the studied CCL5 isolated and that of the GenBank database (NCBI) with 99% identity (443/447) in the forward CCL5. Four mutations were recorded: three transversions - TTT to TTA, AAA to AAT, and GCC to GCA - and one deletion, TGA to TGG. On the other hand, the reverse CCL2 showed 99% identity (436/439) when compared with the GenBank database (NCBI), with three mutations: two deletions - TGA to TG-, and CCA to CC- - and one transversion, ATT to ATA. Also, there was convergence between the studied CXCL10 isolated and that of the GenBank database (NCBI), with 98% identity 64/65 in the forward CXCL10 and one transversion mutation, GGT to GGG.
Conclusions:
New genes for the chemokines CCL2, CCL5 and CXCL10 have been recorded. The results were registered in the NCBI database under accession numbers LC727557, LC727558, and LC727558, respectively. The study revealed the important roles of chemokines in the pathogenesis of MS, suggesting their potential as targets for future therapy.
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