IDENTIFIKASI GEN PENYANDI PROTEIN TRANSPORT SEBAGAI KANDIDAT VAKSIN SUBUNIT TERHADAP BAKTERI Escherichia coli PENYEBAB DIARE WISATAWAN
DOI:
https://doi.org/10.23887/jstundiksha.v9i1.22774Abstrak
Diare masih menjadi masalah global dengan derajat kesakitan dan kematian yang tinggi di berbagai negara. Diare dapat menyerang wisatawan yang mengunjungi objek wisata di kawasan negara berkembang, termasuk Indonesia. Salah satu penyebab diare wisatawan adalah bakteri Escherichia coli pathogen yang memiliki gen penyandi protein transport yang berperan dalam patogenesis diare. Penelitian ini bertujuan untuk mengisolasi dan mengkarakterisasi gen penyandi protein transport sebagai kandidat vaksin subunit terhadap E. coli penyebab diare wisatawan.. Pengambilan sampel feses dilakukan di Ubud Care Clinic pada wisatawan yang melakukan pemeriksaan diare. Sampel dikultur pada media Eosin Metilen Blue, koloni tersangka kemudian diuji biokimia untuk mengkonfirmasi jenis bakteri yang ditemukan. Isolat positif E.coli selanjutnya disubkultur pada media Nutriet Broth, kemudian DNA E.coli diisolasi dengan GeneJET Genomic DNA Purification Kit. DNA hasil isolasi selanjutnya digunakan sebagai template reaksi PCR menggunakan pasangan primer yang mengenali gen aidA penyandi protein transport pada E.coli. Berdasarkan hasil kultur pada media EMB dan hasil uji biokimia pada media TSIA, SIM, Simon Citrat dan uji gula-gula didapatkan sebanyak 20 (59 %) dari 34 sampel feses penderita diare wisatawan positif bakteri E. coli. DNA dari 20 isolat E.coli berhasil diisolasi dengan ukuran sekitar 12000-13000 bp. Hasil PCR dari 20 sampel DNA bakteri E. coli terdapat 5 sampel positif mengandung gen aidA yang ditunjukkan dengan adanya pita DNA tunggal dengan ukuran sekitar 585 bp. Keberadaan gen aidA yang menyandi protein transport menunjukkan sifat patogenitas, yang membedakan antara E.coli pathogen dengan E.coli non-patogenReferensi
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