Open Access
Research (Published online: 17-09-2023)
15. Designing one-step reverse transcriptase loop-mediated isothermal amplification for serotype O foot-and-mouth disease virus detection during the 2022 outbreak in East Java, Indonesia
Eduardus Bimo Aksono, Mirni Lamid, Rimayanti Rimayanti, Iwan Sahrial Hamid, Mustofa Helmi Effendi, Fedik Abdul Rantam, Widjiati Widjiati, Mufasirin Mufasirin, Heni Puspitasari, Munawaroh Fitria, Nur Syamsiatul Fajar, Lucia Tri Suwanti, Nusdianto Nusdianto, Andi Hamim Zaidan, Yuta Kanai, and Teguh Hari Sucipto
Veterinary World, 16(9): 1889-1896

Eduardus Bimo Aksono: Department of Veterinary Medicine, Faculty of Veterinary Medicine, Universitas Airlangga, Surabaya, 60115, Indonesia; Institute of Life Science, Technology and Engineering, Universitas Airlangga, Surabaya, 60115, Indonesia.
Mirni Lamid: Department of Veterinary Medicine, Faculty of Veterinary Medicine, Universitas Airlangga, Surabaya, 60115, Indonesia.
Rimayanti Rimayanti: Department of Veterinary Medicine, Faculty of Veterinary Medicine, Universitas Airlangga, Surabaya, 60115, Indonesia.
Iwan Sahrial Hamid: Department of Veterinary Medicine, Faculty of Veterinary Medicine, Universitas Airlangga, Surabaya, 60115, Indonesia.
Mustofa Helmi Effendi: Department of Veterinary Medicine, Faculty of Veterinary Medicine, Universitas Airlangga, Surabaya, 60115, Indonesia.
Fedik Abdul Rantam: Department of Veterinary Medicine, Faculty of Veterinary Medicine, Universitas Airlangga, Surabaya, 60115, Indonesia.
Widjiati Widjiati: Department of Veterinary Medicine, Faculty of Veterinary Medicine, Universitas Airlangga, Surabaya, 60115, Indonesia.
Mufasirin Mufasirin: Department of Veterinary Medicine, Faculty of Veterinary Medicine, Universitas Airlangga, Surabaya, 60115, Indonesia; Institute of Tropical Disease, Universitas Airlangga, Surabaya, 60115, Indonesia.
Heni Puspitasari: Institute of Tropical Disease, Universitas Airlangga, Surabaya, 60115, Indonesia.
Munawaroh Fitria: Department of Clinical Pathology, Faculty of Medicine, Universitas Airlangga, Surabaya, 60132, Indonesia.
Nur Syamsiatul Fajar: Institute of Tropical Disease, Universitas Airlangga, Surabaya, 60115, Indonesia.
Lucia Tri Suwanti: Department of Veterinary Medicine, Faculty of Veterinary Medicine, Universitas Airlangga, Surabaya, 60115, Indonesia.
Nusdianto Nusdianto: Department of Veterinary Medicine, Faculty of Veterinary Medicine, Universitas Airlangga, Surabaya, 60115, Indonesia.
Andi Hamim Zaidan: Institute of Life Science, Technology and Engineering, Universitas Airlangga, Surabaya, 60115, Indonesia.
Yuta Kanai: Research Institute for Microbial Diseases, Osaka University, Suita, Osaka, 565-0871, Japan.
Teguh Hari Sucipto: Institute of Tropical Disease, Universitas Airlangga, Surabaya, 60115, Indonesia.

doi: 10.14202/vetworld.2023.1889-1896

Article history: Received: 28-03-2023, Accepted: 23-08-2023, Published online: 17-09-2023

Corresponding author: Eduardus Bimo Aksono

E-mail: eduardus-b-a-h@fkh.unair.ac.id

Citation: Aksono EB, Lamid M, Rimayanti R, Hamid IS, Effendi MH, Rantam FA, Widjiati W, Mufasirin M, Puspitasari H, Fitria M, Fajar NS, Suwanti LT, Nusdianto N, Zaidan AH, Kanai Y, and Sucipto TH (2023) Designing one-step reverse transcriptase loop-mediated isothermal amplification for serotype O foot-and-mouth disease virus detection during the 2022 outbreak in East Java, Indonesia, Veterinary World, 16(9): 1889-1896.
Abstract

Background and Aim: Various methods can detect foot-and-mouth disease (FMD) in cows, but they necessitate resources, time, costs, laboratory facilities, and specific clinical specimen submission, often leading to FMD virus (FMDV) diagnosis delays. The 2022 FMD outbreak in East Java, Indonesia, highlighted the need for an easy, inexpensive, rapid, and accurate detection approach. This study aims to devise a one-step reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) technique and phylogenetic analysis to detect the serotype O FMDV outbreak in East Java.

Materials and Methods: Swab samples were collected from the foot vesicles, nasal secretions, and saliva of five suspected FMDV-infected cows in East Java between June and July 2022. The RT-LAMP design used hydroxy naphthol blue dye or SYBR Green I dye, with confirmatory analysis through reverse transcriptase polymerase chain reaction (RT-PCR) targeting 249 base pairs. PCR products underwent purification, sequencing, and nucleotide alignment, followed by phylogenetic analysis.

Results: The RT-LAMP method using hydroxy naphthol blue dye displayed a positive reaction through a color shift from purple to blue in the tube. Naked-eye observation in standard light or ultraviolet (UV) light at 365 nm, with SYBR Green I stain, also revealed color change. Specifically, using SYBR Green I dye, UV light at 365 nm revealed a color shift from yellow to green, signifying a positive reaction. Nucleotide alignment revealed mutations and deletion at the 15th sequence in the JT-INDO-K3 isolate from the East Java FMDV outbreak. Despite differing branches, the phylogenetic tree placed it in the same cluster as serotype O FMDV from Malaysia and Mongolia.

Conclusion: JT-INDO-K3 exhibited distinctions from Indonesian serotype O FMDV isolates and those documented in GenBank. Then, the RT-LAMP method used in this study has a detection limit 10 times higher latter than the conventional RT-PCR limit, without any cross-reactivity among strains.

Keywords: cow, East Java, foot-and-mouth disease virus, reverse transcriptase loop-mediated isothermal amplification, reverse transcriptase polymerase chain reaction, serotype O.