Comparison of two commercial kits and an in-house ELISA for the detection of equine rotavirus in foal feces

Miño, S.; Kern, Angela; Barrandeguy, M.; Parreño, Viviana

doi:10.1016/j.jviromet.2015.05.002

Cited by 18 publications

(9 citation statements)

References 41 publications

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“…Therefore, the commercially available human RVA ELISA assays have been used for detection of RVA in animals (Bailey et al, 2013). However, as a result of antigenic drift, there are several variants or genotypes of RVA VP6 that have been described (Mino et al, 2015). Differences among these VP6 genotypes influence the performance of various commercially available VP6 based immunoassays.…”

Section: Introductionmentioning

confidence: 99%

“…Therefore, diagnostic kit validation for each species is necessary. Not all the assays have been validated for use in all animal species, and this is particularly true for horses (Mino et al, 2015). In fact, one human RVA rapid immunoassay developed for patient-side use was shown not to work in detection of RVA in horses (Slovis et al, 2014).…”

Section: Introductionmentioning

confidence: 99%

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Comparison of electron microscopy, ELISA, real time RT-PCR and insulated isothermal RT-PCR for the detection of Rotavirus group A (RVA) in feces of different animal species

Soltan

Tsai

Lee

et al. 2016

Journal of Virological Methods

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There is no gold standard for detection of Rotavirus Group A (RVA), one of the main causes of diarrhea in neonatal animals. Sensitive and specific real-time RT-PCR (rtRT-PCR) assays are available for RVA but require submission of the clinical samples to diagnostic laboratories. Patient-side immunoassays for RVA protein detection have shown variable results, particularly with samples from unintended species. A sensitive and specific test for detection of RVA on the farm would facilitate rapid management decisions. The insulated isothermal RT-PCR (RT-iiPCR) assay works in a portable machine to allow sensitive and specific on-site testing. The aim of this investigation was to evaluate a commercially available RT-iiPCR assay for RVA detection in feces from different animal species. This assay was compared to an in-house rtRT-PCR assay and a commercially available rtRT-PCR kit, as well as an ELISA and EM for RVA detection. All three PCR assays targeted the well-conserved NSP5 gene. Clinical fecal samples from 108 diarrheic animals (mainly cattle and horses) were tested. The percentage of positive samples by ELISA, EM, in-house rtRT-PCR, commercial rtRT-PCR, and RT-iiPCR was 29.4%, 31%, 36.7%, 51.4%, 56.9%, respectively. The agreement between different assays was high (81.3-100%) in samples containing high viral loads. The sensitivity of the RT-iiPCR assay appeared to be higher than the commercially available rtRT-PCR assay, with a limit of detection (95% confidence index) of 3-4 copies of in vitro transcribed dsRNA. In conclusion, the user-friendly, field-deployable RT-iiPCR system holds substantial promise for on-site detection of RVA.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Comparison of electron microscopy, ELISA, real time RT-PCR and insulated isothermal RT-PCR for the detection of Rotavirus group A (RVA) in feces of different animal species

Soltan

Tsai

Lee

et al. 2016

Journal of Virological Methods

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“…Target rotaviruses in different titers were recognized by the immobilized monoclonal antibodies, resulting in a change in the impedance spectra of immunosensor interface. The limit of detection was 2.3 PFU mL −1 which is comparable to the minimal infective dose (1 PFU) and much lower than the limit of detection of commercially available ELISA kits (10 2 Focus forming units (FFU)/mL) .…”

Section: Introductionmentioning

confidence: 99%

Label‐free Electrochemical Impedance Detection of Rotavirus Based on Immobilized Antibodies on Gold Sononanoparticles

Attar

Mandli

Ennaji³

et al. 2016

Electroanalysis

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1Introduction Rotavirus (RV), an on-enveloped double-stranded RNA (dsRNA) virus of the Reoviridae family discovered in 1973 [1],i so ne of the majorv iral agent associated with infantile diarrhead isease.O bviously,R Vs of species A (RVA )a re recognized as an important cause of severe childhood gastroenteritis worldwide and accounts for 450,000d eaths per year [2,3].Rotaviruses are ubiquitous and globally responsible for an estimated 5% of all deaths in childreny ounger than 5 years of age,m ostly in developing and low-income countries [4].T hey are also considered as zoonotic emerging viruses,w hich can be transmitted among humans and many animal species [5,6].Routine diagnosisf or laboratory detection of rotavirus particles or antigens is oftenb ased on rapidd etection, generally by enzyme immunosorbent assay (EIA), latex agglutination assay [7,8],p olymerase chain reaction (PCR) and quantitative RT-PCR( qRT-PCR) [1].Regardless of individual advantageso ft he most frequentlyu sed conventional methods,s ome limitations and drawbacks are known as laborious,t ime consuming, expensive, and are not amenable for on-site use.T hough, enzyme-linked immunosorbent assay( ELISA) is thep referred sensing system for the revealing rotavirus antigen, it requires expensives pectrophotometric reader that involves chromogenic reagent, preceded by,c onsiderable incubation,w ashing and reaction steps.F urthermore,R T-PCR based assays involve high technological equipment demands,m ore expensivea nd technical expertise,a nd severalsample preparation steps [9][10][11].Rotaviruses are infectious in very low doses of 1p laque forming unit (PFU);t herefore even af ew viral particles in water can infect ap erson [12,13],h ence requiringh ighly sensitive methods for environmentalm onitoring. At present, there are no available commercialized biosensor devices for rotavirus recognition, thus,t he improvement of new diagnostic strategiess uch as biosensors, has ownedaunique place to combat the virus disease.Biosensors,particularly electrochemical immunosensorsb ased on high specificity in antigen/antibody recognition appear very promising for the determinationo fb iomolecules.T hey combinei mmunoassay and biosensor techniques by making ad irect correlation among the concentrationo fa ntigen and the electrochemical analytical signal [14,15].Obviously,e lectrochemical technique is ap romising alternative system and effective candidatef or rapid,o n-site and point-of-care analysis. Hence,i no rder to amplify the signal and improve sensitivity,d ifferent strategies have been focused on usingv arious nanostructures such as noble metal nanoparticles,c arbon nanotubes,p olymeric nanoparticles and graphene [16][17][18][19].Abstract:C urrently,r otaviruses (RVs) are the leading cause of severe childhoodg astroenteritis worldwide.T his study proposes as ensitive and simple label-free assayf or determiningr otaviruses using,f or the first time,e lectrochemical impedance spectroscopy (EIS). Thei mmunosensor is fabricated by self-assembled monolayers (S...

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“…562,611,612 Fecal PCR analysis is also useful in documenting Clostridium, Lawsonia intracellularis, and Cryptosporidium spp., as well as both equine rotavirus and equine coronavirus. 539,616 Serologic methods, evaluating the presence of antibodies, have been used in the diagnosis of Neorickettsia risticii and Lawsonia intracellularis. 539,572,615 A number of commercial assays are also available for the detection of rotavirus antigen.…”

Section: Ancillary Diagnostic Testsmentioning

confidence: 99%

Clinical Approach to Commonly Encountered Problems

Hines¹

2018

Equine Internal Medicine

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Comparison of two commercial kits and an in-house ELISA for the detection of equine rotavirus in foal feces

Cited by 18 publications

References 41 publications

Comparison of electron microscopy, ELISA, real time RT-PCR and insulated isothermal RT-PCR for the detection of Rotavirus group A (RVA) in feces of different animal species

Comparison of electron microscopy, ELISA, real time RT-PCR and insulated isothermal RT-PCR for the detection of Rotavirus group A (RVA) in feces of different animal species

Label‐free Electrochemical Impedance Detection of Rotavirus Based on Immobilized Antibodies on Gold Sononanoparticles

Clinical Approach to Commonly Encountered Problems

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