Pengaruh Enzim Taq Polimerase dan Suhu Annealing terhadap Amplifikasi Gen Tropomyosin Sarcoptes scabiei

Taufik Mulya Perdana, Alfin Harjuno Dwiputro, Yogik Onky Silvana Wijaya, Tri Baskoro Tunggul Satoto

Abstract


Skabies adalah penyakit kulit yang menjadi masalah kesehatan masyarakat, terutama di negara berkembang. Hingga saat ini, belum ada alat bantu diagnostik yang dapat digunakan untuk menunjang diagnosis skabies. Salah satu kandidat alat diagnostik yang gencar dikembangkan adalah uji imonologis yang mendeteksi antibodi anti-protein struktural dari Sarcoptes scabiei seperti tropomyosin. Selama ini, pembuatan tropomyosin rekombinan menggunakan sumber berupa copy DNA (cDNA) yang cenderung mahal. Untuk menekan beban produksi, digunakanlah genomic DNA (gDNA) sebagai sumber materi genetik. Akan tetapi, pembuatan tropomyosin rekombinan dari gDNA belum pernah dikerjakan. Oleh karena itu, dilakukanlah studi optimisasi polymerase chain reaction (PCR) ini sebagai langkah awal pengembangan alat bantu diagnostik tersebut. Penelitian ini bertujuan untuk mengetahui kondisi amplifikasi gen tropomyosin yang optimal dengan mempelajari efek penggunaan MyTaqTM HS Red Mix (MeridianBioscience) and GoTaq® Green Master Mix. (Promega) serta suhu annealing yang berbeda pada amplifikasi gen tropomiosin S. scabiei. Hasil PCR dengan MyTaqTM HS Red Mix pada suhu annealing 57.1°C, 60.9°C, 63.4°C, dan 65°C menghasilkan pita yang terlihat jelas. PCR dengan menggunakan GoTaq® Green Master Mix tidak menghasilkan amplifikasi DNA. Oleh karena itu, amplifikasi gen tropomiosin S. scabiei paling baik dilakukan dengan menggunakan MyTaqTM HS Red Mix, dengan suhu annealing 65°C.

Keywords


sarcoptes scabiei, skabies, optimisasi pcr, tropomiosin

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DOI: https://doi.org/10.24853/jkk.20.2.138-145

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