Probe‐based real‐time reverse transcription loop‐mediated isothermal amplification (RRT‐LAMP) assay for rapid and specific detection of foot‐and‐mouth disease virus

Abstract

Foot-and-mouth disease (FMD) is a highly contagious transboundary disease of domestic and wild cloven-hoofed animals. FMD is associated with considerable economic losses due to decreased animal productivity and disease-associated restrictions on international trade in animal and animal products (Jamal & Belsham, 2013; World Organisation for Animal Heath (OIE), 2017). FMD virus (FMDV) belongs to the genus Aphthovirus within the family Picornaviridae and has been classified into seven genetically and immunologically distinct serotypes, including O, A, C, Southern African Territories (SAT) 1, SAT 2, SAT 3 and Asia 1; these serotypes have undergone further diversification into multiple topotypes and genetic lineages (Knowles & Samuel, 2003). The genetic and immunological diversity of this virus has been recognized as a great challenge with respect to efforts to diagnose and to prevent this disease.

Evolution of the Probe-Based Loop-Mediated Isothermal Amplification (LAMP) Assays in Pathogen Detection

“…Kubota et al developed the assimilating probe-based LAMP and subsequently designed a simple, low-power, portable handheld device to enable real-time detection of the LAMP amplicons [41,51]. This assay was mainly used for singleplex detection of viruses, and the sensitivities of this assay in the detection of foot-and-mouth disease virus (FMDV) and porcine circovirus 3 (PCV3) were 10 2 copies per µL and 50 copies per reaction, respectively [52,53].…”

Advances in the differential molecular diagnosis of vesicular disease pathogens in swine

“…To date, several reverse transcription LAMP (RT-LAMP) assays have been developed that are capable of detecting all seven FMDV serotypes and serotype-specific detection of O, A, C and Asia 1 FMDVs ( Lim et al, 2020 ). Timely diagnosis is essential for the control, monitoring and eradication of FMD.…”

“…They found that lyophilization greatly improved the storage stability of test reagents without jeopardizing the assays' performance, which further supported the potential on field diagnosis of RT-LAMP assay ( Howson et al, 2017a ). Recently, Lim et al developed a probe-based real-time reverse transcription loop-mediated isothermal amplification (RRT-LAMP) assay for rapid and specific detection of FMDV, and the detection limit of the assay was 10 2 copies/μL which is comparable to that achieved by qRT-PCR ( Lim et al, 2020 ). Rapid and accurate detection of SVA is necessary to confirm the disease causing agent, and to initiate the implementation of control processes.…”

“…Then the stem-loop structure containing multiple initiation sites was used as the template of LAMP reaction for cyclic amplification, and many nucleotide chains with different lengths were generated finally. The results of LAMP assay can be visualized through the following ways (Nie, 2005;Almasi et al, 2014;Mansour et al, 2015;Peng et al, 2021;Bhat et al, 2022), ( 1 To date, several reverse transcription LAMP (RT-LAMP) assays have been developed that are capable of detecting all seven FMDV serotypes and serotype-specific detection of O, A, C and Asia 1 FMDVs (Lim et al, 2020). Timely diagnosis is essential for the control, monitoring and eradication of FMD.…”

Diagnosis of Ruminant Viral Diseases with Loop-Mediated Isothermal Amplification

“…Developed test identified all 7 FMDV serotypes while having no cross-reactivity with other tested viruses. In diagnostic samples, RT-LAMP assay had 97.1% agreement with gold-standard RT-qPCR as well [ 79 ]. There were other FMDV RT-LAMP assays designed to identify serotypes circulating in Korea [ 78 ], India [ 80 ], and Middle East [ 81 ].…”