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Title: | Validasi Metode Identifikasi DNA Kerbau (Bubalus bubalis) Menggunakan Real-Time Polymerase Chain Reaction |
Other Titles: | Validation method of the Buffalo (Bubalus bubalis) DNA Identification using Real-time Polymerase Chain Reaction |
Authors: | Latif, Hadri Adnyane, I Ketut Mudite Putri, Ellena Adinda |
Issue Date: | 22-Jul-2021 |
Publisher: | IPB University |
Abstract: | Banyak kasus pemalsuan produk pangan berbahan dasar daging yang
dijumpai di masyarakat. Penambahan daging dengan daging spesies lain yang
tidak terjamin mutunya, biasanya dengan kualitas lebih rendah dapat berdampak
pada ekonomi dan kesehatan. Identifikasi spesies melalui penggandaan molekul
DNA target dalam satu reaksi dapat dilakukan dengan metode real-time PCR.
Penelitian ini bertujuan untuk memvalidasi metode real-time Polymerase Chain
Reaction (qPCR) dalam identifikasi DNA kerbau pada produk pangan asal hewan.
Metode qPCR dengan SYBR green melting curve analysis digunakan sebagai
pengujian validasi metode DNA kerbau dengan menggunakan kit komersial
QIAGEN DNeasy Food Mericon Kit. Parameter validasi metode yang digunakan
dalam penelitian ini meliputi repeatability,reproducibility, limit of detection, dan
spesifisitas. Hasil penelitian pada parameter validasi repeatability yaitu sampel
positif teramplifikasi 100% sedangkan untuk sampel negatif tidak teramplifikasi.
Hasil uji t terhadap nilai Ct dan Tm pada parameter reproducibility DNA kerbau
menunjukkan bahwa tidak terdapat perbedaan yang nyata dalam mendeteksi DNA
kerbau pada dua pengujian dengan menggunakan metode qPCR, nilai limit
deteksi untuk PCR spesifik kerbau sebesar 0,0625%, serta tidak terdapat reaksi
silang antar DNA spesies pada pengujian spesifisitas. Metode qPCR dapat
dijadikan sebagai metode pengujian untuk mengidentifikasi DNA spesies yang
tidak dikehendaki pada pangan asal hewan yang beredar di masyarakat,
khususnya terkait pemalsuan daging. There are many cases of fraud meat-based food products found in society. Meat substitution with an unsecured meat, usually of less quality, could have economic and health repercussions. Identification of species utilizing the multiplication of target DNA molecules in a single reaction can be carried out by real-time PCR. This research was aimed to validate the real-time polymerase chain reaction (qPCR) method for identifying buffalo DNA on animal food product. The qPCR method with SYBR green melting curve analysis was used as a validation test for the buffalo DNA. The validation parameters used in this study were repeatability, reproducibility, limit of detection, and specificity. The results showed that on repeatability validation parameters were 100% on positive and negative samples. The t test results on the Ct and Tm values of buffalo DNA reproducibility parameters showed that there was no significant difference in detecting buffalo DNA in two examination with using the qPCR method. The limit of detection (LOD) for buffalo using this method was 0,0625%, and there was no cross-reaction between DNA species in the specificity test. The qPCR method can be used as a testing method to identify the DNA of unwanted species in food of animal origin circulating in the community, especially related to meat adulteration. |
URI: | http://repository.ipb.ac.id/handle/123456789/107667 |
Appears in Collections: | UT - Animal Disease and Veterinary Health |
Files in This Item:
File | Description | Size | Format | |
---|---|---|---|---|
cover.pdf Restricted Access | Cover | 582.02 kB | Adobe PDF | View/Open |
B04170022_ Ellena Adinda Putri.pdf Restricted Access | Fullteks | 973.31 kB | Adobe PDF | View/Open |
lampiran.pdf Restricted Access | Lampiran | 973.31 kB | Adobe PDF | View/Open |
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