Foot‐and‐mouth disease virus detection on a handheld real‐time polymerase chain reaction platform

Hole, Kate; Nfon, Charles

doi:10.1111/tbed.13227

Cited by 19 publications

(21 citation statements)

References 21 publications

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“…Point-of-care qPCR for viral pathogens can now deliver a result in less than 20 minutes (Melchers et al, 2017). Similarly, several mobile qPCR instruments have been brought to the market and have been successfully deployed to deliver veterinary diagnoses in remote locations on actionable timescales (Hole & Nfon, 2019). However, the cost of such approaches is likely to be prohibitive in respect to their application to the detection of pathogens in non-human subjects.…”

Section: Discussionmentioning

confidence: 99%

In-situnon-lethal rapid test to accurately detect the presence of the nematode parasite,Anguillicoloides crassus, in European eel,Anguilla anguilla

Noia

Poole

Kaufmann

et al. 2020

Preprint

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Anguillicoloides crassus is an invasive nematode parasite of the European eel, Anguilla anguilla, and one of the primary drivers of eel population collapse. The presence of the parasite has been shown to impact many features of eel physiology and life history. Early detection of the parasite is vital to limit the spread of A. crassus. However, until recently, accurate diagnosis of infection could only be achieved via terminal dissection. To support A. anguilla fisheries management in the context of A. crassus we developed a rapid non-lethal and non-invasive environmental DNA method to detect the presence of the parasite in the swim bladder. Screening of 131 wild eels was undertaken between 2017 and 2019 Ireland and UK to validate the procedure. DNA extractions and PCR were conducted using both a Qiagen Stool kit at Glasgow University and in situ using Whatman qualitative filter paper No. 1 and a miniPCR DNA Discovery System™.Primers were specifically designed from the cytochrome oxidase mtDNA gene region. In situ extraction and amplification takes approx. 3h for up to 16 individuals with higher specificity and sensitivity compare to the laboratory Qiagen kit extraction. The local diagnostic procedure demonstrated Positive Predictive Values at 96% and Negative Predictive Values at 87%. Our method will be a powerful tool in the hands of fisheries managers to help protect this iconic but critically endangered species. It will allow a non-invasive monitoring of the A. crassus dispersion across the European waters.

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Section: Discussionmentioning

confidence: 99%

In-situnon-lethal rapid test to accurately detect the presence of the nematode parasite,Anguillicoloides crassus, in European eel,Anguilla anguilla

Noia

Poole

Kaufmann

et al. 2020

Preprint

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“…Liquid exudates from the thawed muscle (MJ) released into the plastic bag were harvested with a sterile pipette and transferred into cryovials. Then, 10% tissue suspensions of the lymph nodes and tonsils were prepared using Precellys Lysing Kits (BER-P000918LYSK0A0, ESBE Scientific, Markham, ON, USA) as previously described [ 27 , 28 ]. MJ and tissue suspensions were stored at −70 °C until they were used for testing.…”

Section: Methodsmentioning

confidence: 99%

“…RNA extractions were performed on MJ, serum, oral swabs, and tissue suspensions using the Applied Biosystems MagMAX-96 Viral RNA Isolation Kit (AMB1836-5, Life Technologies, Burlington, ON, USA) together with a MagMAX Express-96 Deep Well Magnetic Particle Processor (Life Technologies) as described previously [ 28 ].…”

Section: Methodsmentioning

confidence: 99%

Detection of Foot-and-Mouth Disease Virus in Swine Meat Juice

et al. 2020

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Foot-and-mouth disease virus (FMDV) is a highly contagious agent that impacts livestock industries worldwide, leading to significant financial loss. Its impact can be avoided or minimized if the virus is detected early. FMDV detection relies on vesicular fluid, epithelial tags, swabs, serum, and other sample types from live animals. These samples might not always be available, necessitating the use of alternative sample types. Meat juice (MJ), collected after freeze-thaw cycles of skeletal muscle, is a potential sample type for FMDV detection, especially when meat is illegally imported. We have performed experiments to evaluate the suitability of MJ for FMDV detection. MJ was collected from pigs that were experimentally infected with FMDV. Ribonucleic acid (RNA) was extracted from MJ, sera, oral swabs, and lymph nodes from the same animals and tested for FMDV by real-time reverse transcription polymerase chain reaction (rRT-PCR). MJ was also tested for FMDV antigen by Lateral Flow Immunoassay (LFI). FMDV RNA was detected in MJ by rRT-PCR starting at one day post infection (DPI) and as late as 21 DPI. In contrast, FMDV RNA was detected in sera at 1–7 DPI. Antigen was also detected in MJ at 1–9 DPI by LFI. Live virus was not isolated directly from MJ, but was recovered from the viral genome by transfection into susceptible cells. The data show that MJ is a good sample type for FMDV detection.

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“…To overcome such limitation, on-site devices capable of performing FMDV diagnosis in the field have been developed, such as the fully automated cartridge-based RT-qPCR diagnostic system, Enigma MiniLab R (18). Another handheld RT-qPCR device, Biomeme two3 TM (two3) has also been developed and evaluated as a field-deployable platform for FMDV diagnosis, in which the sensitivity was shown to be almost comparable to RT-qPCR using the ABI7500 platform (19). The RNA samples can be extracted with an on-site RNA extraction kit such as Biomeme M1 Sample Prep TM cartridge kit, a method that is dispensable of laboratory equipment and chemicals such as microcentrifuge, alcohol, phenol, and chloroform (19).…”

Section: Description Detection Limit Tested Clinical Samples Referencesmentioning

confidence: 99%

Advances in the Diagnosis of Foot-and-Mouth Disease

et al. 2020

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Foot-and-mouth disease (FMD) is a devastating livestock disease caused by foot-and-mouth disease virus (FMDV). Outbreaks of this disease in a country always result in conspicuous economic losses to livestock industry and subsequently lead to serious socioeconomic damages due to the immediate imposition of trade embargo. Rapid and accurate diagnoses are imperative to control this infectious virus. In the current review, enzyme-linked immunosorbent assay (ELISA)-based methods used in FMD diagnosis are extensively reviewed, particularly the sandwich, liquid-phase blocking, and solid-phase competition ELISA. The differentiation of infected animals from vaccinated animals using ELISA-based methods is also highlighted, in which the role of 3ABC polyprotein as a marker is reviewed intensively. Recently, more studies are focusing on the molecular diagnostic methods, which detect the viral nucleic acids based on reverse transcription-polymerase chain reaction (RT-PCR) and RT-loop-mediated isothermal amplification (RT-LAMP). These methods are generally more sensitive because of their ability to amplify a minute amount of the viral nucleic acids. In this digital era, the RT-PCR and RT-LAMP are progressing toward the mobile versions, aiming for on-site FMDV diagnosis. Apart from RT-PCR and RT-LAMP, another diagnostic assay specifically designed for on-site diagnosis is the lateral flow immunochromatographic test strips. These test strips have some distinct advantages over other diagnostic methods, whereby the assay often does not require the aid of an external device, which greatly lowers the cost per test. In addition, the on-site diagnostic test can be easily performed by untrained personnel including farmers, and the results can be obtained in a few minutes. Lastly, the use of FMDV diagnostic assays for progressive control of the disease is also discussed critically.

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Foot‐and‐mouth disease virus detection on a handheld real‐time polymerase chain reaction platform

Cited by 19 publications

References 21 publications

In-situnon-lethal rapid test to accurately detect the presence of the nematode parasite,Anguillicoloides crassus, in European eel,Anguilla anguilla

In-situnon-lethal rapid test to accurately detect the presence of the nematode parasite,Anguillicoloides crassus, in European eel,Anguilla anguilla

Detection of Foot-and-Mouth Disease Virus in Swine Meat Juice

Advances in the Diagnosis of Foot-and-Mouth Disease

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