Deteksi Gen COL2A1 dan SLITRK1 Menggunakan DNA Preservasi Asal Saliva Manusia yang Diberikan Kombinasi Antimikroba dan Penghambat Dnase

Abstract

Backgrounds. The use of DNA is important in many fields. DNA is often obtained from blood samples. Compared to blood, saliva has several advantages. In saliva, DNase and microorganisms capable of degrading DNA are present. DNA preservation in saliva requires a combination method of antibiotics that can be bactericidal while inhibiting DNase activity. Detection of genes encoding proteins, namely COL2A1 and SLITRK1, can prove the success of the preservation of the human genome from saliva. Aims. This research aims to determine the role of a combination of antibiotics and DNase inhibitors to preserve DNA from human saliva by detecting COL2A1 and SLITRK1 genes. Methods. This was a purely pilot experimental study. Saliva samples were collected from 10 subjects and divided into no-treatment (NT) and treatment (T) groups. The NT group was not given anything. Group T was supplemented with gentamicin sulfate, clindamycin, ketoconazole, and EDTA. Blood samples served as positive controls. Salivary DNA was extracted using the spin column method on days 0, 3, 7, 10, and 14 of storage at room temperature. DNA purity and concentration were measured using a nanophotometer. COL2A1 and SLITRK1 genes were detected using the polymerase chain reaction method and then visualized using 1% agarose gel electrophoresis. Results. The average concentration of DNA obtained from the extraction was 29.40 ng/μL and the average purity was 1.70. The PCR results of the NT group showed more multiple bands, smears, and undetectable genes. Meanwhile, more target gene bands were detected in group T. Conclusions. The use of a combination of antimicrobials and DNase inhibitors can be a method to preserve DNA in closed containers at room temperature for up to 14 days.