Deteksi Gen ESRl, EGFR, PTK2, JAK2, dan FGFR2 pada DNA Preservasi Asal Saliva Manusia yang Diberi Kombinasi Antimikroba dan Penghambat DNase

Abstract

Background: Genomic DNA preservation protects and stores DNA for a long

time in storage containers so that DNA can be used at any time. DNA serves as a carrier of genetic information from one generation to another. DNA identification can be obtained from blood, buccal swabs, saliva and biopsy tissue. Taking DNA from saliva samples is more beneficial because it is non-invasive, does not require skilled personnel and is easy to do anywhere. Saliva contains white blood cells that produce DNA. Saliva also contains enzymes, hormones, immunoglobulins and normal flora such as bacteria, fungi, protozoa, and viruses. Enzymes and oral microorganisms can degrade DNA. The combination of antimicrobials can create sterile saliva, while deoxyribonuclease inhibitors (DNase) help preserve human DNA.

Aims: to preserve the DNA of human salivary origin given a combination of antimicrobial and dnase inhibitors for 28 days stored in a sealed container at room temperature. This study used a purely experimental design.

Methods: Saliva was collected from 10 subjects, divided into the untreated group

(TP) and the treatment group (P) with the addition of 1,500 µg/ml. gentamicin sulfate, 200 µg/mL clindamycin, 1,000 µg/ml. ketoconazole, and I mM EDTA DNA from both groups (TP and P) was extracted by spin column method based on salivary storage days ie. days 0, 7, 14, 21, and 28. The concentration and purity of DNA are measured by spectrophotometric rules. Target genes (ESR1. EGFR, PTK2, JAK2, and FGFR2) were detected by polymerase chain reaction (PCR) and visualized by electrophoresis of 1% agarose gel and ethidium bromide.

Results: PCR group P showed the presence of all target gene bands (ESR1, EGFR, PTK2, JAK2, and FGFR2) at days 0 to 28, while in the TP group, target gene bands disappeared at days 7, 14, 21 and 28. The combination of antimicrobials and DNAse activity inhibitors can preserve human DNA of salivary origin in sealed containers at room temperature for up to 28 days.

Conclusion: The combination of antimicrobe and DNase inhibitors can preserve human DNA of saliva origin stored at room temperature for up to 28 days.