Perubahan profil metagenomik mikrobioma oral dalam saliva pasien positif COVID-19 setelah berkumur obat kumur komersil
L atar Belakang: COVID-19 merupakan pandemi yang disebabkan oleh SARS-CoV-2 dan menjadi salah satu penyakit infeksius dengan kasus aktif dan kasus kematian yang tinggi di dunia. SARS-CoV-2 dapat ditemukan di berbagai tubuh manusia yang terinfeksi salah satunya adalah saliva yang menjadi pusat penyebaran virus melalui droplets maupun aerosol. Beberapa obat kumur komersil seperti Povidone Iodine (PVP-I), Sodium Chlorite (NaClO2), dan Cetylpyridinium Chloride (CPC) memiliki agen antiseptik yang dapat menurunkan viral load, tingkat keparahan penyakit, serta meminimalisir transimisi virus pada pasien positif COVID-19. Tujuan: Mengetahui perubahan profil metagenomik mikrobioma oral dalam saliva pasien positif COVID-19 setelah berkumur obat kumur komersil. Metode: Penelitian eksperimental laboratorik dilakukan dengan mengumpulkan sampel saliva pasien positif COVID-19 baik sebelum maupun sesudah berkumur obat kumur komersil atau air steril. Ekstraksi DNA dilakukan pada seluruh sampel. Setelah ekstraksi DNA selesai, seluruh sampel diamplifikasi pada daerah V3-V4 gen 16S rRNA menggunakan PCR dilanjutkan dengan penambahan index pada amplikon dengan PCR, lalu sampel dimurnikan dan divalidasi. Lalu konsentrasi DNA dinormalisasi sesuai platform Illumina diikuti dengan pooling pada setiap sampelnya. Dilanjutkan dengan metode Next Generation Sequencing dengan parameter 300 siklus dan kontrol spike-in PhiX 20%. Hasil: Profil mikrobioma oral pada saliva pasien positif COVID-19 dari segi richness dan diversity tidak berbeda secara signifikan dibandingkan kontrol negatif. Genus Streptococcus, Prevotella, dan Veillonella ditemukan lebih tinggi jumlahnya pada pasien positif COVID-19 sebelum berkumur obat kumur komersil. Jumlah spesies ditemukan berubah antara sebelum dan sesudah berkumur obat kumur komersil. Kesimpulan: Berkumur obat kumur komersil menurunkan beberapa spesies patogen yang mengakibatkan penyakit baik lokal maupun sistemik tanpa merubah komposisi mikrobioma rongga mulut secara keseluruhan sehingga mendukung mikrobioma oral yang sehat.
B ackground: COVID-19 is a pandemic caused by SARS-CoV-2 and is one of the most infectious diseases with high morbidity and mortality in the world. SARS-CoV-2 can be found in various part of human bodies whose infected, one of which is saliva, which is the main cause of the viral transmission via droplets or aerosols. Several commercial mouthwashes such as Povidone Iodine (PVP-I), Sodium Chlorite (NaClO2), and Cetylpyridinium Chloride (CPC) have antiseptic agents that can reduce viral load, disease severity, and transmission of the virus in positive COVID-19 patients. Objective: To determine changes in the metagenomic profile of the oral microbiome in saliva of COVID-19 positive patients after gargling commercial mouthwash. Methods: Experimental laboratory research was first conducted by collecting each saliva samples from COVID-19 positive patients both before and after gargling commercial mouthwash or sterile water. DNA extraction was performed on all samples. Then, the DNA were amplified in the V3-V4 regions of 16S rRNA gene using PCR followed by addition of index to the amplicon with PCR, the samples were purified and validated. Afterwards, the DNA concentration was normalized according to the Illumina platform followed by pooling on each sample. Followed by sequencing using Next Generation Sequencing (NGS) method with sequencing parameter of 300 cycles and PhiX control spike-in of 20%. Results: The oral microbiome profile in the saliva of COVID-19 positive patients in terms of richness and diversity was not significantly different from negative control. At the genus level, the relative abundance of Streptococcus, Prevotella, and Veillonella was higher in oral microbiota of positive COVID-19 patients before gargling commercial mouthwash. The number of species found differed before and after gargling commercial mouthwash. Conclusion: Gargling commercial mouthwash reduce several pathogenic species that cause both local and systemic disease without changing the composition of the overall oral microbiome thereby supporting and maintaining a healthy oral microbiome.