Development and evaluation of one-step real-time RT-PCR assay for improved detection of foot-and-mouth disease virus serotypes circulating in Egypt

El-Bagoury, Gabr; El-Habashy, Rawan A; Ayman, H Mahmoud; Naglaa, M Hagag; Zowalaty, Mohamed E. El

doi:10.1016/j.jviromet.2022.114525

Cited by 9 publications

(3 citation statements)

References 40 publications

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“…The detailed process of RT-qPCR for FMDV detection was carried out according to Bagoury. 28 In addition, this method and RT-qPCR were used to detect a variety of common animal viruses, such as FMDV, SFV, ASFV, LSDV, and PRV. Finally, 18 samples containing FMDV positive and negative samples were used to verify the potential application of the method in real samples.…”

Section: Detection Of Fmdv By Combining Cha and Hcrmentioning

confidence: 99%

Enzyme-free and sensitive method for single-stranded nucleic acid detection based on CHA and HCR

Chen¹,

Huang²,

Nie³

et al. 2023

Anal. Methods

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Section: Detection Of Fmdv By Combining Cha and Hcrmentioning

confidence: 99%

Enzyme-free and sensitive method for single-stranded nucleic acid detection based on CHA and HCR

Chen¹,

Huang²,

Nie³

et al. 2023

Anal. Methods

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“…Vaccination for one serotype offers limited or no protection against others, emphasizing the need to identify the correct vaccine for epidemic control [4,12,13]. Yoon et al [14] outlined conventional FMDV detection methods, including antigen-specific enzyme-linked immunosorbent assay, cytopathic effect observation in cultures [3], reverse transcriptase polymerase chain reaction (RT-PCR) [15], and real-time quantitative polymerase chain reaction (qPCR) [16,17]. These tests demand resources, time, costs, laboratory facilities, and special clinical specimen delivery, leading to FMDV diagnosis delays [18].…”

Section: Introductionmentioning

confidence: 99%

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

Aksono,

Lamid,

Rimayanti

et al. 2023

Vet World

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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.

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“…Likewise, Bachanek-Bankowska et al were able to discern FMDV A, O, SAT 1 and 2, restricted to viruses found in East Africa ( 27 ). El Bagoury et al ( 28 ) produced rRT-PCR assays capable of detecting and distinguishing O and SAT3 viruses circulating in Egypt. Several other lineage-specific FMDV rRT-PCR assays have been reported ( 25 , 29 – 32 ).…”

Section: Introductionmentioning

confidence: 99%

Development of reverse-transcriptase, real-time PCR assays to distinguish the Southern African Territories (SAT) serotypes 1 and 3 and topotype VII of SAT2 of Foot-and-Mouth Disease Virus

Chestley

Sroga²,

Nebroski³

et al. 2022

Front. Vet. Sci.

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Foot-and-Mouth Disease Virus (FMDV), the causative agent of Foot-and-Mouth Disease, is a highly feared, economically devastating transboundary pathogen. This is due to the virus' extremely contagious nature and its ability to utilize multiple transmission routes. As such, rapid and accurate diagnostic testing is imperative to the control of FMD. Identification of the FMDV serotype is necessary as it provides the foundation for appropriate vaccine selection and aids in outbreak source tracing. With the vast genetic diversity, there is a desperate need to be able to characterize FMDV without relying on prior knowledge of viral serotypes. In this study, the Neptune bioinformatics tool was used to identify genetic signatures specific to each Southern African Territories (SAT) 1, 2 and 3 genomes but exclusionary to the other circulating FMDV serotypes (A, O, Asia1, and the heterologous SAT1, SAT2 and/or SAT3). Identification of these unique genomic regions allowed the design of TaqMan-based real-time reverse transcriptase PCR (rRT-PCR) primer/probe sets for SAT1, SAT2 and SAT3 viruses. These assays were optimized using prototypic FMDV cell culture isolates using the same reagents and thermocycling conditions as the FMDV pan-serotype 3D rRT-PCR assay. Cross-reactivity was evaluated in tandem with the FMDV pan-serotype 3D rRT-PCR utilizing representative strains from FMDV serotypes A, O, Asia1, SAT1, SAT2 and SAT3. The SAT1, SAT2, and SAT3 primer/probe sets were specific for the homologous serotype and exclusionary to all others. SAT1 and SAT3 primer/probe sets were able to detect several topotypes, whereas the SAT2 assay was revealed to be specific for topotype VII. The SAT2 topotype VII specificity was possibly due to the use of sequence data deposited post-2011to design the rRT-PCR primers and probes. Each assay was tested against a panel of 99 bovine tissue samples from Nigeria, where SAT2 topotype VII viruses were correctly identified and no cross-reactivity was exhibited by the SAT1 and 3 assays. These novel SAT1, SAT3 and SAT2 topotype VII rRT-PCR assays have the potential to detect and differentiate circulating FMD SAT viruses.

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Development and evaluation of one-step real-time RT-PCR assay for improved detection of foot-and-mouth disease virus serotypes circulating in Egypt

Cited by 9 publications

References 40 publications

Enzyme-free and sensitive method for single-stranded nucleic acid detection based on CHA and HCR

Enzyme-free and sensitive method for single-stranded nucleic acid detection based on CHA and HCR

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

Development of reverse-transcriptase, real-time PCR assays to distinguish the Southern African Territories (SAT) serotypes 1 and 3 and topotype VII of SAT2 of Foot-and-Mouth Disease Virus

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