IRVAN MAULANA, . (2022) VALIDASI FORMULA KIT DETEKSI BAKTERI PATOGEN Salmonella typhi DAN Salmonella typhimurium PENYEBAB KERACUNAN PANGAN DENGAN REAL TIME POLYMERASE CHAIN REACTION. Sarjana thesis, UNIVERSITAS NEGERI JAKARTA.
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Abstract
Kasus keracunan pangan masih sering terjadi dan merupakan salah satu penyebab kematian dan kesakitan di Indonesia. Pada tahun 2019, keracunan pangan di indonesia terdapat 77 kasus (BPOM, 2019). Bakteri Salmonella typhi dan Salmonella typhimurium merupakan bakteri gram negatif yang menyebabkan keracunan pangan. Metode deteksi yang cepat, akurat, dan spesifik sangat dibutuhkan untuk mendeteksi penyebab keracunan pangan. Pada penelitian sebelumnya telah didapatkan kondisi dan formula optimum dalam mendeteksi bakteri tersebut dengan menggunakan Real Time PCR. Hasil penelitian sebelumnya dijadikan landasan untuk pengembangan prototype kit deteksi. Pada penelitian ini dilakukan validasi yang bertujuan untuk mengkonfirmasi hasil penelitian sebelumnya sehingga didapatkan produk yang reprodusibel dan dilakukan optimasi konsentrasi primer dan waktu pra-denaturasi. Hasil penelitian ini menunjukkan bahwa primer fim-C bakteri Salmonella typhi dan primer pef bakteri Salmonella typhimurium teramplifikasi pada 95 basepair (bp) dan 139 bp dengan suhu annealing 60ºC dan standar konsentrasi DNA 50 ng/μL. Hasil validasi uji konfirmasi Real Time PCR bakteri Salmonella typhi dan Salmonella typhimurium pada pra-denaturasi 3 menit dengan konsentrasi 10 pmol didapatkan nilai Ct sesuai standar dengan penelitian sebelumnya. Nilai Ct yang didapatkan sebesar 13,96 untuk bakteri S. typhi dan Ct 15,64 untuk bakteri S. typhimurium. Hasil uji konfirmasi sampel pangan menunjukkan hasil positif dibandingkan dengan standar SNI bakteri S. typhi pada sampel pangan susu fresh, susu basi, kaldu jamur fresh, kaldu jamur basi kaldu daging fresh dan kaldu daging basi. Pada bakteri S. typhimurium sampel pangan susu fresh susu basi, kaldu jamur fresh, kaldu jamur basi, kaldu daging fresh dan kaldu daging basi menunjukkan hasil positif dibandingkan dengan standar SNI mengandung bakteri S. typhimurium. Berdasarkan hasil yang diperoleh dapat dikatakan bahwa tahap validasi kondisi terhadap kultur murni bakteri berhasil dilakukan dengan menghasilkan data yang konsisten dan bersifat reprodusibel. ************************************************************ Cases of food poisoning are still common and are one of the causes of death and death in Indonesia. In 2019, there were 77 cases of food poisoning in Indonesia (BPOM, 2019). Salmonella typhi and Salmonella typhimurium are gram-negative bacteria that cause food poisoning. A fast, accurate, and specific detection method is needed to detect poisoning. In previous studies, the optimum conditions and formulas for detecting these bacteria have been obtained using Real Time PCR. The results of previous studies are used as the basis for the development of a prototype detection kit. In this study, validation was carried out, which aims to confirm the results of previous studies in order to obtain a reproducible product and optimize the primary concentration and pre-denaturation time. The results of this study showed that fim-C primers for Salmonella typhi and pef primers for Salmonella typhimurium were amplified at 95 base pairs (bp) and 139 bp with an annealing temperature of 60ºC and a standard DNA concentration of 50 ng/μL. The results of the Real Time PCR confirmation test of Salmonella typhi and Salmonella typhimurium bacteria at a pre-denaturation of 3 minutes with a concentration of 10 pmol obtained the Ct value according to the standard with previous studies. The Ct value obtained was 13.96 for S. typhi bacteria and Ct 15.64 for S. typhimurium bacteria. The results of the confirmation test of food samples showed positive results compared to the SNI standard for S. typhi bacteria on food samples of fresh milk, stale milk, fresh mushroom broth, stale mushroom broth, fresh meat broth, and stale meat broth. The bacteria S. typhimurium food samples of fresh milk, fresh mushroom broth, stale mushroom broth, fresh meat broth, and meat broth showed positive results compared to the SNI standard containing S. typhimurium bacteria. Based on the results obtained, it can be said that the condition validation stage for pure cultures was successfully carried out by producing consistent and reproducible data.
| Item Type: | Thesis (Sarjana) |
|---|---|
| Additional Information: | 1). Prof. Dr. Muktiningsih Nurjayadi M.Si.; 2). Vira Saamia S.Si, M.Biomed |
| Subjects: | Sains > Kimia |
| Divisions: | FMIPA > S1 Kimia |
| Depositing User: | sawung yudo |
| Date Deposited: | 07 Nov 2024 03:32 |
| Last Modified: | 07 Nov 2024 03:32 |
| URI: | http://repository.unj.ac.id/id/eprint/51961 |
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