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All Journal HAYATI Journal of Biosciences VALENSI Tadris: Jurnal keguruan dan Ilmu Tarbiyah Indonesian Journal of Biotechnology Science and Technology Indonesia Jurnal Penelitian Pendidikan IPA (JPPIPA) Jurnal SOLMA Jurnal Riset Pendidikan Kimia (JRPK) Jurnal Riset Sains dan Kimia Terapan JSMARTech : Journal of Smart Bioprospecting and Technology Makara Journal of Science SEGARA : Jurnal Pengabdian Kepada Masyarakat Jurnal Kimia Unand Jurnal Bioteknologi & Biosains Indonesia (JBBI) Jurnal Bioteknologi & Biosains Indonesia
Kurniadewi, Fera
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Agriculture, Biological Sciences & Forestry Humanities Biochemistry, Genetics & Molecular Biology Chemical Engineering, Chemistry & Bioengineering Chemistry Dentistry Earth & Planetary Sciences Education Energy Environmental Science Health Professions Immunology & microbiology Materials Science & Nanotechnology Medicine & Pharmacology Neuroscience Physics 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 Achmad Ridwan, Achmad Ade Danova Agus Setiawan Akbar, Dandy Allanas, Edith Amyyana, Afwu Hayyi Angelina, Helzi Anggraeni, Rosita Gio Anis, Mhd. Aqilah, Amadita Shafa Asworo, Yuliana Dwi Aulia Rachma, Lu'luunnisa Azizah, Puan Aqila Azzahra, Maharanianska Barkah, Eka Berkahingrum, Ayu Bunga Falana, Isfasona Chavasiri, Warinthorn Chen, Shyi-Tien Dalia Sukmawati Danova, Ade Darmiyanti, Waskitarini Darwis, Zulmanelis Dewanti, Farradilla Alenia Didin Mujahidin El Enshasy, Hesham El-Enshasy, Hesham Ali Elenshasy, Hesham Ali Enshasy, Hesham Ali El Fahriza, Tiara Fitria Handayani Fitriyanti, Anisa Ghufrooni, Rifqi Grace, Grace Hadi Nugrahaningsih, Tritiyatma Hadinugrahaningsih, Tritiyatma Hermawati, Elvira Iqbal Musthapa Irwanto Irwanto Istantia Chantika, Kinanti Jefferson Lynford Declan Juliansyah, Dandy Akbar Kartika, Irma Kartika, Irma Ratna Kartikawati, Nindya Galuh Krisdawati, Ismaya Laurensia, Laurensia Maria Paristiowati Maulana, Irvan Muktiningsih Nurjayadi Musie, Royna Rahma Nugroho, Aidahaya Hanannisa Nurhayadi, Muktiningsih Nurul Fadillah Octaviany, Sheila Putri Fajriani, Fathya Putri, Adinda Myra Amalia Putri, Gladys Indira Putri, Gusti Angieta Rahmawati, Atikah Nur Ratna Kartika, Irma Rosita Rosita Rustini, Yeyet Saamia, Vira Saamia4, Vira Saputra, Irwan Saputro, Dwi Anna Oktaviani Sari, Ely Puspita Septiani, Elsa Shangkara, Muhammad Arkent Sihombing, Ananda Indah Putri Siti Fatimah SRI RAHAYU Susilo, Ahmad Dwi Syafriani, Yolanda Febrica Umiatin, Umiatin Widiawan, Ages Kayla Shyfa Wiranatha, I Made Wonganan, Piyanuch Yuli Rahmawati