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All Journal HAYATI Journal of Biosciences VALENSI Microbiology Indonesia Acta Biochimica Indonesiana Sarwahita : Jurnal Pengabdian Kepada Masyarakat Jurnal Riset Sains dan Kimia Terapan JSMARTech : Journal of Smart Bioprospecting and Technology Prosiding Seminar Nasional Pengabdian Kepada Masyarakat Makara Journal of Science Jurnal Bioteknologi & Biosains Indonesia (JBBI) Jurnal Kesehatan Masyarakat Jurnal Bioteknologi & Biosains Indonesia
Jefferson Lynford Declan
(1) Department Of Chemistry, Faculty Of Mathematics And Natural Science, Universitas Negeri Jakarta, Gedung KH. Hasjim Asj'ari, 6th Floor, Jl. Rawamangun Muka, Jakarta Timur, 13220, Indonesia (2) Research Center For Detection Of Pathogenic Bacteria,
Agriculture, Biological Sciences & Forestry Humanities Biochemistry, Genetics & Molecular Biology Chemistry Earth & Planetary Sciences Economics, Econometrics & Finance Education Energy Environmental Science Health Professions Immunology & microbiology Materials Science & Nanotechnology Medicine & Pharmacology Neuroscience Public Health Social Sciences Other

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Journal : Makara Journal of Science

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Detection Method for Escherichia coli Using Real-Time Polymerase Chain Reaction Targeting the yhaV Gene Nurjayadi, Muktiningsih; Fitriyanti, Anisa; Musie, Royna Rahma; Putri, Gusti Angieta; Azizah, Puan Aqila; Angelina, Helzi; Grace, Grace; Sihombing, Ananda Indah Putri; Setiawan, Agus; Declan, Jefferson Lynford; Putri, Gladys Indira; Juliansyah, Dandy Akbar; Fatimah, Siti; Berkahingrum, Ayu; Kartika, Irma Ratna; Kurniadewi, Fera; Saamia, Vira; Chen, Shyi-Tien; Aboemolak, Bassam; Enshasy, Hesham Ali El
Makara Journal of Science Vol. 29, No. 3
Publisher : UI Scholars Hub

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Abstract

Escherichia coli is a foodborne pathogenic bacterium that can cause diarrhea, while yhaV is a virulence-associated gene linked to the toxin–antitoxin system in E. coli. This study was aimed at evaluating the confirmation, specificity, and sensitivity of a yhaV gene primer using real-time polymerase chain reaction. The yhaV-targeting PCR successfully amplified a DNA fragment with an amplicon length of 207 bp (base pairs) under an annealing temperature optimized to a range of 54 °C to 62 °C via gradient PCR. The PCR using the primer pair produced a consistent Ct (cycle threshold) of 14.14 ± 0.05 and showed a single peak in the melting curve at a Tm (melting temperature) of 83.67 °C ± 0.02. The specificity test indicated that the yhaV primer effectively distinguished E. coli from nontarget bacteria on the basis of differences in Ct and Tm values. The sensitivity analysis showed that the PCR directed toward the primer pair successfully detected E. coli at a minimum concentration of 2.24 pg/µL, with a Ct value of 29.93 and a detection limit of 31.5 × 102 CFU. These results suggest that yhaV-based real-time PCR quickly and accurately identifies E. coli. Primer designs that target yhaV have the potential to be developed as components of a rapid, specific, and sensitive kit for detecting E. coli in food samples.
Co-Authors Aboemolak, Bassam Abomoelak, Bassam Abumoelak, Bassam Adinda Myra Amalia Putri Adinda Myra Amalia Putri Agus Setiawan AGUS SETIAWAN Akbar, Dandy Ananda Indah Putri Sihombing Angelina, Helzi Anggraeni, Rosita Gio Anisa Fitriyanti Annisa Maharani Athiyah Layla Atikah Nur Rahmawati Ayu Berkahingrum Azizah, Puan Aqila Azzahra, Maharanianska Bassam Abomoelak Bassam Abomoelak Berkahingrum, Ayu Chen, Shyi-Tien Dalia Sukmawati Dandy Akbar Juliansyah Dandy Akbar Juliansyah Dandy Akbar Juliansyah Dwi Ana Oktaviani El Enshasy, Hesham El-Enshasy, Hesham Ali Elenshasy, Hesham Ali Enshasy, Hesham Ali El Fahriza, Tiara Fera Kurniadewi Fera Kurniadewi Fitriyanti, Anisa Gladys Indira Putri Gladys Indira Putri Gladys Indira Putri Grace Grace, Grace Gusti Angieta Putri Hairishah, Nazrisya Helzi Angelina Hesham A. El Enshasy Hesham Ali Al-Enshashy Hesham Ali El-Enshasy I Made Wiranatha I Made Wiranatha Irma Ratna Kartika Irvan Maulana Irvan Maulana Irwan Saputra Ismaya Krisdawati Ismaya Krisdawati Ismaya Krisdawati Juliansyah, Dandy Akbar Kartika, Irma Ratna Krisdawati, Ismaya Kurniadewi, Fera Maharani Azka Azzahra Maharanianska Azzahra Maulana, Irvan Mellyna Fitriani Muktiningsih Nurjayadi Musie, Royna Rahma Nabilla Anisa Putrie Nasywa Fhelia Salta Nida Nur Afifah NOVITASARI Pardiana Puan Aqila Azizah Putri, Adinda Myra Amalia Putri, Gladys Indira Putri, Gusti Angieta Rahmawati, Atikah Nur Ririn Gustini Rizkahana Syehfia Rosita Gio Anggraeni Royna Rahma Musie Saamia, Vira Saamia4, Vira Saputro, Dwi Anna Oktaviani Sarah Adilisa Kartini Shangkara, Muhammad Arkent Shyi-Tien Chen Sihombing, Ananda Indah Putri Sintia Mardita Siti Fatimah SITI FATIMAH SRI RAHAYU Sri Rahayu Tiara Fahriza Uswatul Nisa Vira Saamia Vira Saamia Vira Saamia Wiranatha, I Made