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All Journal HAYATI Journal of Biosciences Jurnal Riset Sains dan Kimia Terapan JSMARTech : Journal of Smart Bioprospecting and Technology Jurnal Kesehatan Masyarakat
Abomoelak, Bassam
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Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Chemistry Earth & Planetary Sciences Education Materials Science & Nanotechnology Medicine & Pharmacology Neuroscience Public Health

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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.
Detection of the Yersinia enterocolitica Bacteria Targeting the myfA and ystA Genes in Contaminated Vegetable Samples using Real-Time PCR to Develop Rapid Detection of Food Poisoning Bacteria Nurjayadi, Muktiningsih; Anggraeni, Rosita GIo; Putri, Gladys Indira; Declan, Jefferson Lynford; Juliansyah, Dandy Akbar; Fahriza, Tiara; Putri, Adinda Myra Amalia; Berkahingrum, Ayu; Rahmawati, Atikah Nur; Kartika, Irma Ratna; Kurniadewi, Fera; Sukmawati, Dalia; Rahayu, Sri; Saamia, Vira; Wiranatha, I Made; Abomoelak, Bassam; El-Enshasy, Hesham Ali
HAYATI Journal of Biosciences Vol. 32 No. 4 (2025): July 2025
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.4308/hjb.32.4.989-1002

Abstract

Yersinia enterocolitica is a pathogenic bacterium with the ability to survive and multiply in food in a low-temperature environment that can cause death in humans. In previous studies, the optimum annealing temperature of ymoA, ystA, and ail gene primers with amplicons of 185 bp, 123 bp, and 192 bp, respectively, was successfully found. This study aims to develop a pathogenic bacteria detection kit with confirmation, sensitivity, and specificity of myfA and ystA primers in detecting Yersinia enterocolitica bacteria quickly and accurately using the real-time Polymerase Chain Reaction method. The results showed that myfA and ystA primers have optimum annealing temperatures at 60°C with amplicon lengths of 181 bp and 123 bp, respectively. Primer myfA was able to amplify the target with real-time PCR at Ct 12.07±1 and Tm 81±1°C, while the ystA primer at Ct 12.38±1 and Tm 83±1°C. myfA and ystA primers were also able to distinguish target and non-target bacteria based on Ct or Tm. The designed primers successfully detected Yersinia enterocolitica bacteria with the smallest concentration of 0.000439 ng/µL equivalent to 7.024 × 102 CFU. The detection limit obtained is smaller than the contamination threshold set by the Food and Drug Administration (BPOM). Primer myfA and ystA Yersinia enterocolitica also successfully detected the target bacteria in cabbage and lettuce samples artificially. Based on these results, myfA and ystA primers successfully detected Yersinia enterocolitica in vegetable samples using real-time PCR quickly, sensitively, specifically, and accurately.
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.
hpmA Gene as a Detection Method of Proteus mirabilis Bacteria using real-time Polymerase Chain Reaction Nurjayadi, Muktiningsih; Putri, Gladys Indira; Fitriyanti, Anisa; Declan, Jefferson Lynford; Novitasari; Abomoelak, Bassam
Jurnal Kesehatan Masyarakat Vol. 21 No. 2 (2025)
Publisher : Universitas Negeri Semarang in collaboration with Ikatan Ahli Kesehatan Masyarakat Indonesia (IAKMI Tingkat Pusat) and Jejaring Nasional Pendidikan Kesehatan (JNPK)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15294/kemas.v21i2.16214

Abstract

Proteus mirabilis is pathogenic bacteria that can cause gastrointestinal infections, bacteremia, and Urinary Tract Infections (UTI). Therefore, it is necessary to have a fast, sensitive, specific, and accurate detection method to identify Proteus mirabilis. This study aims to determine the confirmation, specificity, and sensitivity test of the hpmA gene primer to detect Proteus mirabilis quick and precise using the real-time Polymerase Chain Reaction method. Gradient Polymerase Chain Reaction results showed the hpmA primer has an amplicons length of 195 bp and the optimum annealing temperature at 60°C. The primer pair produced Ct value of 10.40±0.18 and showed one peak in the melting curve with Tm value of 81.84°C±0.02 by real-time PCR. Furthermore, the hpmA primer was also able to distinguish target and non-target bacteria based on the difference in Ct and Tm value formed. Based on these results, the concentration of bacterial DNA that can be detected by primers reached 3.2 pg/μL, equivalent to the concentration of target bacteria that can be detected by primers is 10.24×102 CFU. In the next step, hmpA primer will be developed to detect Proteus mirabilis in artificial contaminated samples using real-time PCR.
Co-Authors Anggraeni, Rosita Gio Azzahra, Maharanianska Berkahingrum, Ayu Dalia Sukmawati El Enshasy, Hesham El-Enshasy, Hesham Ali Elenshasy, Hesham Ali Fahriza, Tiara Fitriyanti, Anisa Jefferson Lynford Declan Juliansyah, Dandy Akbar Kartika, Irma Ratna Krisdawati, Ismaya Kurniadewi, Fera Maulana, Irvan Muktiningsih Nurjayadi Musie, Royna Rahma NOVITASARI Putri, Adinda Myra Amalia Putri, Gladys Indira Putri, Gusti Angieta Rahmawati, Atikah Nur Saamia, Vira SRI RAHAYU Wiranatha, I Made