Real-time RPA assay for rapid detection and differentiation of wild-type pseudorabies and gE-deleted vaccine viruses

Wang, Jianchang; Liu, Libing; Wang, Jinfeng; Pang, Xiao‐Yu; Yuan, Wanzhe

doi:10.1016/j.ab.2017.12.012

Cited by 19 publications

(10 citation statements)

References 18 publications

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“…The single-tube multiplex RPA can be performed using single or multiple nucleic acid templates. For the former, Kim et al 122 performed a duplex detection of Campylobacter species: Campylobacter coli and Campylobacter jejuni (detection limits of each: 1 CFU mL −1 in pure culture and in chicken broth samples without enrichment; 3 log CFU g −1 in non-enriched egg and chicken meat samples; 1 CFU g −1 in enriched egg and chicken meat samples), and Wang et al 187 conducted a duplex assay to differentiate wild-type and gEdeleted vaccine pseudorabies (limit of detection of 100 viral DNA copies each). However, the highest level of multiplexing is a triplex assay demonstrated by Piepenburg and co-workers for the detection of MRSA I, II and III genotypes (detection limits are 10 genomic copies each).…”

Section: Multiplex Rpamentioning

confidence: 99%

Review: a comprehensive summary of a decade development of the recombinase polymerase amplification

Macdonald

Stetten

2019

Analyst

426

365

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Section: Multiplex Rpamentioning

confidence: 99%

Review: a comprehensive summary of a decade development of the recombinase polymerase amplification

Macdonald

Stetten

2019

Analyst

426

365

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“…El RPA ha sido ampliamente usado para la detección de numerosos patógenos en humanos y animales (Bonney et al, 2017;Boyle et al, 2013;Gao, Jiang, Wang y Wei, 2018;Kim y Lee, 2017;Law et al, 2018;Wang, Liu, Wang, Pang y Yuan, 2018), y más recientemente para enfermedades en plantas, incluyendo la detección de begomovirus (Londoño et al, 2016) igura 1. Alineamiento de la cápside proteica de diferentes especies de begomovirus detectadas en soja y poroto.…”

Section: Resultados Y Discusiónunclassified

Evaluación de la técnica de amplificación por recombinasa y polimerasa (RPA) para la detección de begomovirus presentes en cultivos de soja y poroto en Argentina

Reyna

Pardina²

2018

AgriS

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<p>Los métodos tradicionales de diagnóstico son imprecisos para la identificación de begomovirus. Actualmente se usan técnicas moleculares, que requieren equipamientos sofisticados o procedimientos complejos. La técnica de amplificación mediante recombinasa y polimerasa (RPA) es similar a la reacción en cadena de la polimerasa (PCR), sensible, específica, pero opera a temperatura constante. Con el fin de evaluar la utilización de esta técnica para la detección de begomovirus presentes en soja y poroto en Argentina, se diseñaron iniciadores específicos. Se probaron inicialmente por PCR, con clones de virus detectados en el país, y se logró amplificar una banda de 371pb. La RPA se llevó a cabo con el Twist Amp® Basic kit utilizando muestras de soja y poroto, sanas y enfermas y malezas infectadas. Se probaron dos métodos de conservación de muestras: hojas liofilizadas y hojas mantenidas a -70 ºC; y dos de obtención de ADN: CTAB y molido de la muestra en 0,5M OHNa. La reacción se incubó a 37 ºC durante 30 min, y luego a 65 ºC durante 10 min. Se visualizaron bandas del tamaño esperado en plantas infectadas y no en testigos sanos. No hubo diferencias según los métodos de conservación de material ni con los de extracción de ADN utilizados. Se logró ajustar la RPA para la detección de begomovirus en cultivos de soja y poroto de Argentina.</p>

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“…Dual real-time recombinase polymerase amplification (RPA) assays rapidly identify wild-type PRV and gE-deleted vaccine strains. Specific primers and probes are designed for the conserved regions of the genome (such as gB, gE, gD genes), which can specifically detect and identify wild-type strains and gE-deficient vaccine strains ( Wernike et al, 2011 ; Wang et al, 2018a ). An EvaGreen-based modified multiplex real-time polymerase chain reaction (EGRT-PCR) assay that successfully differentiates wild-type and gE-deficient BoHV-1 strains based on gene-specific melting temperature (Tm) peaks ( Pawar et al, 2017 ).…”

Section: Deleted Marker Vaccines Designed With Gementioning

confidence: 99%

Alphaherpesvirus glycoprotein E: A review of its interactions with other proteins of the virus and its application in vaccinology

et al. 2022

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The viral envelope glycoprotein E (gE) is required for cell-to-cell transmission, anterograde and retrograde neurotransmission, and immune evasion of alphaherpesviruses. gE can also interact with other proteins of the virus and perform various functions in the virus life cycle. In addition, the gE gene is often the target gene for the construction of gene-deleted attenuated marker vaccines. In recent years, new progress has been made in the research and vaccine application of gE with other proteins of the virus. This article reviews the structure of gE, the relationship between gE and other proteins of the virus, and the application of gE in vaccinology, which provides useful information for further research on gE.

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Real-time RPA assay for rapid detection and differentiation of wild-type pseudorabies and gE-deleted vaccine viruses

Cited by 19 publications

References 18 publications

Review: a comprehensive summary of a decade development of the recombinase polymerase amplification

Review: a comprehensive summary of a decade development of the recombinase polymerase amplification

Evaluación de la técnica de amplificación por recombinasa y polimerasa (RPA) para la detección de begomovirus presentes en cultivos de soja y poroto en Argentina

Alphaherpesvirus glycoprotein E: A review of its interactions with other proteins of the virus and its application in vaccinology

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