KUANTIFIKASI HASIL EKSTRAKSI GEN SEBAGAI FAKTOR KRITIS UNTUK KEBERHASILAN PEMERIKSAAN RT PCR

Authors

  • Eka Pratiwi Pusat Penelitian dan Pengembangan Biomedis dan Teknologi Dasar Kesehatan. Kementerian Kesehatan Republik Indonesia, Indonesia
  • Lovendo Ilham Widodo Fakultas Biologi, Universitas Jenderal Soedirman, Indonesia

DOI:

https://doi.org/10.24269/ijhs.v4i1.2293

Abstract

Infeksi virus dan bakteri menjadi penyebab terbanyak infeksi saluran pernafasan di negara berkembang. Influenza Like Illness (ILI) merupakan kelompok penyakit yang komplek dan heterogen yang disebabkan oleh berbagai etiologi. Etiologi ILI terdiri dari kurang lebih 300 jenis virus, bakteri, riketsia dan jamur. Deteksi penyebab kasus ILI saat ini mengalami beberapa kesulitan yaitu dalam menentukan etiologi ILI dari spesimen. Tahap awal dari metode Polymerase Chain Reaction (PCR) dimulai dengan melakukan ekstraksi, yaitu  memisahkan asam nukleat dari komponen sel lainnya, sehingga asam nukleat yang diperoleh dapat dianalisis atau dimodifikasi lebih lanjut dengan teknik biologi molekular lainnya. Setelah ekstraksi, dilakukan kuantifikasi meliputi pengukuran konsentrasi asam nukleat menggunakan alat Qubit fluorometric didapatkan beberapa sampel  RNA nya < 20 ng/mL sehingga tidak dapat terukur alat ini dan hasil pengukuran DNA nya bervariasi dengan range 4,99 – 99,2 ng/mL. Alat  Nanovue Plus juga digunakan untuk pengukuran konsentrasi asam nukleat dan kemurnian, untuk nilai RNA didapatkan range 0,0564 – 0,1396ng/mL lalu konsentrasi DNA berada di kisaran 0,0875 – 0,1365 ng/mL. Pada pengukuran kemurnian menggunakan Nanovue Plus tidak didapatkan DNA yang murni dengan nilai >1,80 dan pengukuran RNA didapat beberapa sampel yang mempunyai kemurnian dengan nilai >2,00.

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Published

2020-03-12

Issue

Vol. 4 No. 1 (2020): March

Section

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