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Development of Bacillus subtilis Detection Method Targeting Genes codY, narH, and ureC Using Polymerase Chain Reaction Nurjayadi, Muktiningsih; Berkahingrum, Ayu; Putri, Adinda Myra Amalia; Rahmawati, Atikah Nur; Anggraeni, Rosita Gio; Fahriza, Tiara; Declan, Jefferson Lynford; Putri, Gladys Indira; Krisdawati, Ismaya; Juliansyah, Dandy Akbar; Azzahra, Maharanianska; Maulana, Irvan; Kartika, Irma Ratna; Kurniadewi, Fera; Sukmawati, Dalia; Rahayu, Sri; Saamia, Vira; Wiranatha, I Made; Abomoelak, Bassam; El Enshasy, Hesham
JSMARTech: Journal of Smart Bioprospecting and Technology Vol. 5 No. 1 (2024): JSMARTech Volume 5, No. 1, 2024
Publisher : JSMARTech

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21776/ub.jsmartech.2024.005.01.26

Abstract

Bacillus subtilis, a causative agent of foodborne illness in bread, cakes, cereals, and cheese leading to symptoms like diarrhea and nausea, was investigated for its role in nosocomial infections, including endocarditis, bacteremia, septicemia, and meningitis. The research focused on three genes: codY, narH, and ureC, which respectively contribute to bacterial survival through biofilm and spore formation, acid resistance, and adaptation to anaerobic conditions. The objective of this study was to evaluate the codY, narH, and ureC genes using the Gradient PCR method for Bacillus subtilis detection in bread and cheese through Real-Time PCR. During the research, bacterial growth was observed with an OD600 of 1.438 in Tryptic Soy Broth (TSB), and colonies of medium size, smooth, cream-coloured, and round shape were successfully isolated on Tryptic Soy Agar (TSA). The DNA template used had a 71.5 ng/µL concentration with an A260/280 ratio of approximately 1.830. The annealing temperature for Gradient PCR used was 53-62°C. The primers successfully amplified codY (175 bp), narH (222 bp), and ureC (153 bp) gene amplicons. The optimal annealing temperature for the primers used was 60°C, as indicated by the presence of a single bright band in electrophoresis. Using these three different genes, testing can also be conducted with the Multiplex PCR method. The next step involves developing a detection kit using optimized primers and annealing temperatures for the identification of Bacillus subtilis in bread and cheese samples using Real-Time PCR.
DISEMINASI PEMBELAJARAN STRUKTUR DAN FUNGSI BIOMOLEKUL PROTEIN BAGI GURU WILAYAH MGMP-1 JAKARTA TIMUR Nurjayadi, Muktiningsih; Saputra, Irwan; Putri, Gladys Indira; Declan, Jefferson Lynford; Krisdawati, Ismaya; Juliansyah, Dandy Akbar; Hairishah, Nazrisya
Bahasa Indonesia Vol 20 No 01 (2023): Sarwahita : Jurnal Pengabdian kepada Masyarakat
Publisher : Lembaga Penelitian dan Pengabdian Kepada Masyarakat

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21009/sarwahita.201.3

Abstract

Materi biokimia di SMA dipelajari pada kelas XII dengan salah satu pembahasan mengenai biomolekul protein. Pada pandemi COVID-19, terjadi perubahan sistem pembelajaran yang membutuhkan berbagai inovasi agar pembelajaran berlangsung efektif. Program pengabdian pada masyakarat ini, melakukan pengembangan pembelajaran struktur dan fungsi biomolekul protein bagi guru MGMP Wilayah Jakarta Timur-1 melalui desiminiasi elektonik modul Struktur dan Fungsi Protein hasil penelitian sebelumnya serta praktikum berbasis bahan rumahan dengan teknik blended learning. Pelaksanaan P2M yang dilakukan bersama Tim UNJ pada Guru-guru Kimia MGMP Jakarta Timur wilayah 1 berjalan dengan lancar. Peserta sangat antusias mengikuti materi yang disampaikan dan ikut berkontribusi dalam pelaksanaan dan mengamati praktikum uji kualitatif asam amino dan protein menggunakan bahan-bahan yang ada disekitar rumah, ramah lingkungan, dan sekaligus menerapkan konsep green chemistry. Berdasarkan hasil umpan balik, 90% peserta merasa puas dengan penyampaian materi yang disampaikan, memperoleh wawasan baru, relevan dengan yang diharapkan, dapat diimplementasikan di sekolah, dan menimbulkan ide baru untuk mendesain pembelajaran yang inovatif. Sehingga diharapkan kegiatan pengabdian kepada masyarakat yang dilakukan oleh dosen Universitas Negeri Jakarta bekerjasama dengan MGMP wilayah Jakarta Timur 1 sebagai wilayah binaan yang meliputi kecamatan Cakung, Duren Sawit, Jatinegara, Matraman, dan Pulogadung, dapat bermanfaat dalam menyebarluaskan IPTEKS dan meningkatkan kompetensi guru.
Deteksi Vibrio parahaemolyticus Menggunakan Primer Gen toxR2 dengan Gradient Polymerase Chain Reaction Nurjayadi, Muktiningsih; Krisdawati, Ismaya; Declan, Jefferson Lynford; Putri, Gladys Indira; Juliansyah, Dandy Akbar; Rahmawati, Atikah Nur; Fitriyanti, Anisa; Musie, Royna Rahma; Kurniadewi, Fera; Sukmawati, Dalia; Saamia, Vira; Wiranatha, I Made; Abomoelak, Bassam; Elenshasy, Hesham Ali
Jurnal Riset Sains dan Kimia Terapan Vol. 11 No. 1 (2025): Jurnal Riset Sains dan Kimia Terapan, Volume 11 Nomor 1, Juni 2025
Publisher : Program Studi Kimia Universitas Negeri Jakarta

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21009/JRSKT.111.03

Abstract

Makanan adalah kebutuhan vital, dengan kriteria utama adalah keamanan, kualitas, dan nilai gizi. Untuk memastikan keamanan makanan, diperlukan metode deteksi yang cepat dan akurat, terutama untuk mendeteksi bakteri patogen penyebab keracunan makanan. Vibrio parahaemolyticus merupakan bakteri patogen yang banyak ditemukan pada makanan laut. Penelitian ini bertujuan untuk mengembangkan metode deteksi cepat V. parahaemolyticus dengan menargetkan gen toxR2 menggunakan Gradient Polymerase Chain Reaction. Gen toxR dipilih karena fungsinya sebagai pengatur penting gen virulensi. Tahapan yang dilakukan meliputi desain primer, penyiapan sampel bakteri dari biakan murni, dan uji amplifikasi menggunakan PCR Gradien. Hasil uji amplifikasi menunjukan bahwa pasangan primer toxR2 berhasil mengamplifikasi pada suhu 58-62°C dengan dihasilkan pita berukuran 137 bp. Berdasarkan hasil tersebut, dapat disimpulkan bahwa PCR Gradien dengan primer toxR2 dapat diaplikasikan untuk mengembangkan alat deteksi V. parahaemolyticus dengan dilakukan uji lanjutan seperti uji konfirmasi, uji spesifisitas, uji sensitivitas, dan uji pada pangan menggunakan Real-Time Polymerase Chain Reaction.
Real-time PCR-based Detection of Foodborne Pathogen Cronobacter sakazakii DNA in Infant Formula Milk with Specific Targeting on the hfq Gene Nurjayadi, Muktiningsih; Juliansyah, Dandy Akbar; Declan, Jefferson Lynford; Putri, Gladys Indira; Krisdawati, Ismaya; Rahmawati, Atikah Nur; Azzahra, Maharanianska; Maulana, Irvan; Putri, Gusti Angieta; Kurniadewi, Fera; Kartika, Irma Ratna; Saamia, Vira; Wiranatha, I Made; Abomoelak, Bassam; El-Enshasy, Hesham Ali
HAYATI Journal of Biosciences Vol. 32 No. 6 (2025): November 2025
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.4308/hjb.32.6.1597-1607

Abstract

Cronobacter sakazakii has been linked to cause meningitis, necrotizing enterocolitis, and sepsis in infants and newborns, with case fatality rates ranging from 40 to 80%. The most common source of infection has been identified as Cronobacter sakazakii-contaminated infant formula. With a relatively specific target hfq gene, this study aims to develop a real-time PCR method to identify Cronobacter sakazakii in infant formula milk. Real-time PCR is used as a detection method because rt- PCR has higher specificity and sensitivity compared to conventional PCR methods. The real-time PCR method also has a higher level of effectiveness and time efficiency compared to conventional PCR. Cronobacter sakazakii ATCC 29544 genomic DNA was isolated and used in a real-time PCR assay. Cronobacter sakazakii DNA was amplified using a primer targeting the hfq gene, yielding a 145 bp amplicon. The results of the real-time PCR test showed that Cronobacter sakazakii DNA with a concentration of 53 ng/µL could be amplified by the primer pairs of hfq gene with Ct values of 11 respectively then had Tm values of 81.7°C±0.5. The specificity test showed that the hfq primer pairs could differentiate between the target and some non-target bacteria. The sensitivity test showed the ability of the primer to detect the smallest concentration of 3.392 pg/µL with a Ct of 26.16. Based on the results obtained, it can be concluded that the hfq primer has the potential to be used as a fast detection method for Cronobacter sakazakii bacteria in infant formula using real-time PCR.