Lestari, Lilis Mustikaning and Dr. Ir. Joni Kusnadi,, M.Si (2022) Optimasi Teknik Direct-PCR Menggunakan Buffer Lisis Untuk Deteksi Adulterasi Daging Kuda (Equus caballus) Pada Produk Pangan Dengan Metode Quantitative Real-Time Polymerase Chain Reaction. Sarjana thesis, Universitas Brawijaya.
Abstract
Pendahuluan: Seiring peningkatan pengetahuan tentang gizi pangan, konsumsi produk berbahan dasar daging semakin meningkat. Hal ini menjadi tuntutan untuk terus selektif dalam memilih makanan. Hingga saat ini, banyak isu mengenai pemalsuan daging pada produk olahan seperti sosis, bakso, kornet dan lainnya demi keuntungan ekonomis. Salah satu isu yang sedang ramai yaitu pemalsuan daging sapi dengan daging kuda di Amerika dan Eropa. Meskipun kuda merupakan hewan halal bagi umat Islam, namun beberapa negara menganggap bahwa kuda merupakan hewan suci yang tidak untuk dikonsumsi. Selain itu, pemalsuan merupakan perbuatan yang tidak dibenarkan karena akan merugikan orang lain. Tujuan: Penelitian ini bertujuan untuk mengoptimasi teknik Direct-PCR dapat diaplikasikan untuk deteksi adulterasi produk pangan sosis dan daging burger dengan menggunakan metode Real-Time PCR dan primer spesifik CO1. Metode: Kinerja buffer lisis diuji pada suhu 75°C, 85°C, dan 95°C dan waktu 5 menit, 15 menit, dan 25 menit. Kemudian dilakukan uji validasi antara lain spesifisitas, sensitivitas dan uji sampling produk komersial. Data akhir dibahas dengan metode deskriptif melalui pendekatan kuantitatif. Hasil: Hasil penelitian menunjukkan bahwa teknik Direct-PCR untuk deteksi adulterasi pangan hewan kuda optimal pada suhu 75°C dan waktu 5 menit. Konsentrasi DNA yang dihasilkan sebesar 339,96 ng/ μl dengan nilai kemurnian DNA 1,51. Pada uji spesifisitas menunjukkan primer CO1 spesifik pada kuda (Equus caballus) dengan sensitivitas hingga 10-3 ng/μl pada Real-Time PCR. Hasil uji sampling pada produk komersial sosis dan daging burger dengan masing-masing tiga merek label daging sapi, terdapat satu sampel yang terdeteksi adanya daging kuda. Aplikasi: Teknik Direct-PCR yang teroptimalisasi dapat digunakan untuk deteksi DNA kuda pada produk olahan daging sosis dan daging burger dengan Real-Time PCR.
English Abstract
Introduction: Along with increasing knowledge about food nutrition, consumption of meat-based products is increasing. This is a demand to continue to be selective in choosing food. Until now, there are many issues regarding adulteration of meat in processed products such as sausages, meatballs, corned beef and others for economic gain. One issue that is currently buzzing is the adulteration of beef with horse meat in America and Europe. Although horses are halal animals for Muslims, some countries consider horses to be sacred animals that are not for consumption. In addition, counterfeiting is an act that is not justified because it will harm other people. Objective: This study aims to optimize the Direct-PCR technique that can be applied to detect adulteration of sausage and patty food products using the Real-Time PCR method and CO1-specific primers. Methods: The performance of the lysis buffer was tested at 75°C, 85°C, and 95°C and a time of 5 minutes, 15 minutes, and 25 minutes. Then, validation tests were carried out including specificity, sensitivity and sampling tests for commercial products. The final data were discussed using a descriptive method through a quantitative approach. Results: The results showed that the Direct-PCR technique for detecting horse food adulteration was optimal at a temperature of 75°C and a time of 5 minutes. The resulting DNA concentration was 339.96 ng/μl with a DNA purity value of 1.51. The specificity test showed a horse-specific CO1 primer (Equus caballus) with a sensitivity of up to 10-3 ng/μl in Real-Time PCR. In the sampling test on sausage and patty commercial products with three brands of beef labels each, there was one sample that detected the presence of horse meat. Application: The optimized Direct-PCR technique can be used for the detection of horse DNA in processed sausage and patty meat products with Real-Time PCR.
| Item Type: | Thesis (Sarjana) |
|---|---|
| Identification Number: | 0522100246 |
| Uncontrolled Keywords: | Adulterasi pangan, Kuda, Real-Time PCR, Teknik Direct-PCR,Direct-PCR Technique, Food Adulteration, Horse, Real-Time PCR |
| Subjects: | 600 Technology (Applied sciences) > 630 Agriculture and related technologies |
| Divisions: | Fakultas Teknologi Pertanian > Keteknikan Pertanian |
| Depositing User: | soegeng sugeng |
| Date Deposited: | 07 Nov 2022 02:09 |
| Last Modified: | 07 Nov 2022 02:09 |
| URI: | http://repository.ub.ac.id/id/eprint/196153 |
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