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Abstrak :
Metode Taqman MGB real time PCR yang cepat merupakan kunci pengawasan pemalsuan daging yang efektif. Penelitian bertujuan mengevaluasi kuantitas, kualitas DNA produk olahan daging babi, serta kandungan DNA babi produk olahan daging sapi yang diduga mengandung babi menggunakan Taqman MGB real time PCR untuk memverifikasi label. Lima produk olahan daging babi, 30 produk olahan daging sapi: dendeng, abon, baso, dan daging asap sebagai sampel, serta daging babi segar sebagai kontrol positif diekstraksi, diukur konsentrasi, kemurnian DNA, dielektroforesis serta diamplifikasi dengan realtime PCR. Konsentrasi, kemurnian DNA, nilai Ct sampel diuji ANAVA satu arah dilanjutkan uji Tukey, kecuali nilai Ct produk olahan daging sapi. Integritas DNA genomnya dianalisis deskriptif. Hasil uji ANAVA menunjukkan ada pengaruh nyata (P˂0,05) konsentrasi, kemurnian DNA dan nilai Ct. Hasil uji Tukey produk olahan daging babi: ada beda nyata konsentrasi DNA sampel dan kontrol positif (P˂0,05), kecuali kornet (P˃0,05). Kemurnian DNA baso dan daging asap berbeda nyata (P˂0,05) dengan kontrol positif. Nilai Ct sampel dan kontrol positif berbeda nyata (P˂0,05), kecuali dendeng (P˃0,05). Hasil uji Tukey produk olahan daging sapi: konsentrasi DNA baso dan daging asap berbeda nyata (P<0,05) dengan kontrol positif, kemurnian DNA kornet berbeda nyata (P<0,05) dengan kontrol positif. Semua DNA genom sampel terfragmentasi ukuran terendahnya sekitar 250 bp dimiliki kornet dan abon. Produk olahan daging dapat meningkat kuantitas DNAnya dan menurun kualitas DNAnya tergantung pada suhu dan bahan tambahan yang diberikan. Tiga puluh produk olahan daging sapi tidak mengandung DNA babi menggunakan Taqman real time PCR yang sensitif dan cepat serta terverifikasi mematuhi peraturan label. ......The fast Taqman MGB qPCR method is key to effective meat adulteration surveillance. This research aimed to evaluate the quantity, quality of DNA from processed pork products and the content of pork DNA in processed beef products suspected of containing pork DNA using the Taqman MGB qPCR to verify labels. Five processed pork products, 30 processed beef products: corned, jerky, shredded, meatballs, and smoked meat were used as samples as well as and fresh pork as a positive control were extracted, DNA concentration and purity were measured, electrophoresed, and amplified with qPCR. The DNA concentration, purity, and Ct value were tested by one-way ANOVA followed by the Tukey test, except for the Ct value of processed beef products. The genomic DNA integrity was analyzed descriptively. The ANOVA showed a significant effect (P˂0.05) on the concentration and purity of DNA and Ct value. Tukey test results for processed pork products: there was a significant difference (P˂0.05) in the DNA concentration of the samples and positive controls, except for corned (P˃0.05). The DNA purity of pork meatballs and smoked pork was significantly different (P˂0.05) from the positive control. The Ct values of the samples and positive control were significantly different (P˂0.05), except for jerky (P˃0.05). The results of the Tukey test for processed beef products: the DNA concentration of beef meatballs and smoked beef was significantly different (P<0.05) with the positive control, and the DNA purity of corned beef was significantly different (P<0,05) with positive control. All genomic DNA samples were fragmented with the smallest size of about 250 bp experienced by corned and shredded. Processed meat products can increase the quantity of DNA and decrease the quality depending on temperature and additives. Thirty processed beef products did not contain pork DNA using the sensitive and fast Taqman qPCR and verified to comply with label regulations.

Abstrak :
Kasus Tuberkulosis (TB) ekstra-paru, yaitu infeksi akibat bakteri Mycobacterium tuberculosis (Mtb) di luar jaringan paru-paru, telah mencakup 15% dari total populasi kasus secara global. Salah satu kasus TBEP yang mendominasi, terletak pada organ gastrointestinal (TBGI). Salah satu penyebab rendahnya laju diagnosis TB ekstra-paru disebabkan oleh beberapa tantangan saat identifikasi. Konfirmasi melalui metode bakteriologis menggunakan uji molekuler quantitative polymerase chain reaction (qPCR) dapat menjadi salah satu pendekatan dalam pengembangan alat uji diagnostik pendukung yang cepat. Tujuan penelitian ini adalah melakukan deteksi gen Insertion Sequence (IS) 6110 menggunakan metode qPCR, dan mengevaluasi nilai diagnostik qPCR IS6110 sebagai biomarka dalam diagnosis TBEP. Penelitian ini menetapkan 103 sampel jaringan biopsi berdasarkan gejala klinis menyerupai TBGI. Metode penelitian terdiri dari isolasi, kuantifikasi, dan amplifikasi DNA menggunakan kit TaqMan qPCR, serta melakukan analisis uji diagnostik berupa sensitivitas, spesifisitas, Positive Predictive Value (PPV), dan Negative Predictive Value (NPV). Keseluruhan kemurnian (A260/280) DNA memiliki nilai rerata yang baik yaitu 1,931. Hasil perhitungan uji diagnostik menunjukkan nilai sensitivitas 58,33% (14/24), spesifisitas 54,43% (43/79), dengan PPV 28%, dan NPV 81%. Penetapan nilai Limit of Detection (LOD) pada TaqMan qPCR mampu mendeteksi hingga 1,4 copies/μL. Berdasarkan hasil penelitian, maka uji TaqMan qPCR dapat menjadi salah satu alat uji diagnosis pendukung pada penanganan TBGI. Akan tetapi, penelitian lebih lanjut menggunakan teknik sekuensing disarankan sebagai bentuk klarifikasi bahwa amplifikasi merupakan gen target yang diharapkan. ......Extra-pulmonary tuberculosis (EPTB), or Mycobacterium tuberculosis (Mtb) infection outside of the lung tissue, accounts for 15% of all tuberculosis cases worldwide. One of the most prevalent EPTB cases is gastrointestinal TB (GITB). The difficulty in identifying EPTB is one indicator of the diagnosis, especially in GITB cases. Therefore, confirmation through bacteriological methods with quantitative molecular polymerase chain reaction (qPCR) techniques can be one approach to developing rapid diagnostic test tools. This study aimed to detect the Insertion Sequence (IS) 6110 gene using the qPCR method and to evaluate the qPCR diagnostic testing to determine its validity in detecting the IS6110 as a TBEP biomarker gene. This study determined 103 biopsy samples based on the GITB clinical inclusion criteria. The research method consisted of isolation, DNA quantification, and DNA amplification with the TaqMan qPCR kit were utilized, followed by diagnostic testing (Sensitivity, Specificity, PPV, and NPV). DNA samples' overall purity (A260/280) had a good average value of 1,931. The results showed that TaqMan qPCR had a sensitivity of 58.33% (14/24), specificity reached 54.43% (43/79), with PPV 28%, and NPV 15,38%. The Limit of Detection (LOD) value in TaqMan qPCR may detect up to 1.4 copies/reaction. Based on the findings of this study, it can be concluded that the qPCR test method can be an option in supporting diagnostic test tools in the treatment of GITB. However, further research using sequencing techniques is suggested as a form of clarification that amplification is the expected gene target.