Development of Reverse Transcription-PCR (Oligonucleotide Probing) Enzyme-Linked Immunosorbent Assays for Diagnosis and Preliminary Typing of Foot-and-Mouth Disease: a New System Using  Simple and Aqueous-Phase Hybridization

A reverse transcription-PCR (RT-PCR)–enzyme-linked immunosorbent assay system that detects a relatively conserved region within the RNA genome of all seven serotypes of foot-and-mouth disease virus (FMDV) has been developed. The high specificity of the assay is achieved by including a rapid hybridization step with a biotin-labeled internal oligonucleotide. The assay is highly sensitive, fast, and easy to perform. A similar assay, based on a highly variable region of the FMDV genome and employing a single asymmetric RT-PCR and multiple hybridization oligonucleotides, was developed to demonstrate the method's ability to type FMDV. Based on our theoretical and practical knowledge of the methodology, we predict that similar assays are applicable to diagnosis and strain differentiation in any system amenable to PCR amplification.

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Reverse transcription-PCR using a Primer Set Targeting the 3D Region Detects Foot-and-Mouth Disease Virus with High Sensitivity

10.1101/305086 ◽

2018 ◽

Author(s):

Tatsuya Nishi ◽

Toru Kanno ◽

Nobuaki Shimada ◽

Kazuki Morioka ◽

Makoto Yamakawa ◽

...

Keyword(s):

Reverse Transcription ◽

Foot And Mouth Disease ◽

Mouth Disease ◽

Clinical Samples ◽

Rt Pcr ◽

Pcr Assay ◽

Reverse Transcription Pcr ◽

Mouth Disease Virus ◽

Foot And Mouth ◽

Pcr Assays

AbstractBecause foot-and-mouth disease (FMD) has the potential to spread extensively, methods used for its diagnosis must be rapid and accurate. Therefore, reverse transcription-PCR (RT-PCR) plays an important diagnostic role. Here we designed the primer set FM8/9 to amplify 644 bases of the conserved 3D region of all seven serotypes of FMD virus (FMDV). We compared the performance of RT-PCR assays using FM8/9 with that using the primer set 1F/R targeting the 5’-UTR described in the manual of the World Organization for Animal Health. The detection limits of the RT-PCR assays were determined for 14 strains representing all serotypes. Compared with the sensitivities of the RT-PCR assay using 1F/R, those using FM8/9 were 101-to 104-fold higher for eight strains. To assess the validity of the methods for analyzing clinical samples, sera and saliva samples from pigs and cows infected with FMDV were collected daily and analyzed using the two PCR assays. The FM8/9 assay detected FMDV from all infected pigs and cows for longer times compared with the 1F/R assay, therefore revealing higher sensitivity for the clinical samples. Our results suggest that the FM8/9 RT-PCR assay is highly sensitive and is therefore suitable for the diagnosis of FMD.

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Comparison of stabilisers for development of a lyophilised multiplex reverse-transcription PCR mixture for rapid detection of foot and mouth disease virus serotypes

Revue Scientifique et Technique de l OIE ◽

10.20506/rst.33.3.2323 ◽

2014 ◽

Vol 33 (3) ◽

pp. 859-867 ◽

Cited By ~ 1

Author(s):

G.K. SHARMA ◽

S. MAHAJAN ◽

B.B. DAS ◽

R. RANJAN ◽

A.N. KANANI ◽

...

Keyword(s):

Disease Virus ◽

Reverse Transcription ◽

Rapid Detection ◽

Foot And Mouth Disease ◽

Mouth Disease ◽

Reverse Transcription Pcr ◽

Mouth Disease Virus ◽

Foot And Mouth

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Robust Real-Time Reverse Transcription-PCR for Detection of Foot-and-Mouth Disease Virus Neutralizing Carryover Contamination

Journal of Clinical Microbiology ◽

10.1128/jcm.01944-15 ◽

2015 ◽

Vol 54 (1) ◽

pp. 216-219 ◽

Cited By ~ 1

Author(s):

Ji-Hyeon Hwang ◽

Yong-Keol Shin ◽

So-Yeon Park ◽

Jeesoo Kim ◽

Su-Mi Kim ◽

...

Keyword(s):

Real Time ◽

Reverse Transcription ◽

Diagnostic Method ◽

Foot And Mouth Disease ◽

Mouth Disease ◽

Reverse Transcription Pcr ◽

Outbreak Period ◽

Mouth Disease Virus ◽

Foot And Mouth ◽

Specific Restriction

During an outbreak of foot-and-mouth disease (FMD), real-time reverse transcription-PCR (rRT-PCR) is the most commonly used diagnostic method to detect viral RNA. However, while this assay is often conducted during the outbreak period, there is an inevitable risk of carryover contamination. This study shows that the carryover contamination can be prevented by the use of target-specific restriction endonuclease in that assay.

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Reverse transcription‐PCR using a primer set targeting the 3D region detects foot‐and‐mouth disease virus with high sensitivity

Transboundary and Emerging Diseases ◽

10.1111/tbed.13202 ◽

2019 ◽

Cited By ~ 1

Author(s):

Tatsuya Nishi ◽

Toru Kanno ◽

Nobuaki Shimada ◽

Kazuki Morioka ◽

Makoto Yamakawa ◽

...

Keyword(s):

Disease Virus ◽

Reverse Transcription ◽

Foot And Mouth Disease ◽

High Sensitivity ◽

Mouth Disease ◽

Reverse Transcription Pcr ◽

Mouth Disease Virus ◽

Foot And Mouth

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Detection of foot-and-mouth disease virus from culture and clinical samples by reverse transcription-PCR coupled to restriction enzyme and sequence analysis

Veterinary Research ◽

10.1051/vetres:2002059 ◽

2003 ◽

Vol 34 (1) ◽

pp. 105-117 ◽

Cited By ~ 24

Author(s):

Margarita S�iz ◽

Diana B. de la Morena ◽

Esther Blanco ◽

Jos� I. N��ez ◽

Rufino Fern�ndez ◽

...

Keyword(s):

Sequence Analysis ◽

Disease Virus ◽

Restriction Enzyme ◽

Reverse Transcription ◽

Foot And Mouth Disease ◽

Mouth Disease ◽

Clinical Samples ◽

Reverse Transcription Pcr ◽

Mouth Disease Virus ◽

Foot And Mouth

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Validation of reverse transcriptase RT-PCR for determining the concentration of 146S particles of foot-and-mouth disease virus in the raw material for vaccines

Veterinary Medicine Journal ◽

10.30896/0042-4846.2018.21.9.58-63 ◽

2018 ◽

Vol 21 (9) ◽

pp. 58-63

Author(s):

D.A. Lozovoy ◽

◽

М.I. Doronin ◽

V.А. Starikov ◽

M.N. Guseva ◽

...

Keyword(s):

Reverse Transcriptase ◽

Disease Virus ◽

Foot And Mouth Disease ◽

Mouth Disease ◽

Raw Material ◽

Rt Pcr ◽

Mouth Disease Virus ◽

Foot And Mouth

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Use of a Standardized Bovine Serum Panel To Evaluate a Multiplexed Nonstructural Protein Antibody Assay for Serological Surveillance of Foot-and-Mouth Disease

Clinical and Vaccine Immunology ◽

10.1128/cvi.00227-07 ◽

2007 ◽

Vol 14 (11) ◽

pp. 1472-1482 ◽

Cited By ~ 11

Author(s):

Julie Perkins ◽

Satya Parida ◽

Alfonso Clavijo

Keyword(s):

Nonstructural Protein ◽

Foot And Mouth Disease ◽

Mouth Disease ◽

Enzyme Linked Immunosorbent Assay ◽

Reference Laboratory ◽

Additional Information ◽

Mouth Disease Virus ◽

Foot And Mouth ◽

Antibody Assays ◽

Serological Surveillance

ABSTRACT Liquid array technology has previously been used to show proof of principle of a multiplexed nonstructural protein serological assay to differentiate foot-and-mouth disease virus-infected and vaccinated animals. The current multiplexed assay consists of synthetically produced peptide signatures 3A, 3B, and 3D and the recombinant protein signature 3ABC in combination with four controls. To determine the diagnostic specificity of each signature in the multiplex, the assay was evaluated against a naive population (n = 104) and a vaccinated population (n = 94). Subsequently, the multiplexed assay was assessed by using a panel of bovine sera generated by the World Reference Laboratory for foot-and-mouth disease in Pirbright, United Kingdom. This serum panel has been used to assess the performance of other singleplex enzyme-linked immunosorbent assay (ELISA)-based nonstructural protein antibody assays. The 3ABC signature in the multiplexed assay showed performance comparable to that of a commercially available nonstructural protein 3ABC ELISA (Cedi test), and additional information pertaining to the relative diagnostic sensitivity of each signature in the multiplex was acquired in one experiment. The encouraging results of the evaluation of the multiplexed assay against a panel of diagnostically relevant samples promote further assay development and optimization to generate an assay for routine use in foot-and-mouth disease serological surveillance.

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