Rapid detection of sacbrood virus (SBV) by one-step reverse transcription loop-mediated isothermal amplification assay

Yang, Jing; Yang, Ruifu; KeFei, Shen; Xiang-wei, Peng; Xiong, Tao; Liu, Zuohua

doi:10.1186/1743-422x-9-47

Cited by 5 publications

(1 citation statement)

References 4 publications

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“…Another TaqMan method designed for Chinese SBV isolates from North China [ 14 ] does not exactly match (two mismatched nucleotides in both the primers and one in the probe) with our Fujian clone and Yunnan Ac SBV collected from East and Southwest China, respectively. Other published methods with SYBR Green qPCR [ 15 ] and loop-mediated PCR [ 16 , 17 ] also have a few mismatched nucleotides. These findings surprised us when we attempt to adapt these methods in our studies, and these mismatched nucleotides may lead to false-negative results.…”

Section: Introductionmentioning

confidence: 99%

Updating Sacbrood Virus Quantification PCR Method Using a TaqMan-MGB Probe

Huang

Zhang

Mehmood

et al. 2021

Veterinary Sciences

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Sacbrood virus (SBV) is a common honey bee virus disease. SBV variants and strains identified in Asian honey bees, Apis cerana, have created confusion in identifications. Although the regional names indicated the expansions of the virus in new regions, pathogenesis, and genomes of these variants are not distinct enough to be a separate virus species. However, current SBV qPCR methods may not detect newly identified A. cerana SBV variants (Ac SBV) according to the genome sequences. Since these Ac SBV can naturally infect A. mellifera and possibly other hymenopterans, ignorance of Ac SBV variants in detection methods is simply unwise. In this report, we updated the qPCR method based on Blanchard’s design that used conserved regions of VP1 to design a TaqMan method with an MGB (minor groove binder) probe. We tested the method in bees and hornets, including A. mellifera, A. cerana, and Vespa velutina. The updated primers and the probe can match published SBV and Ac SBV genomes in databases, and this updated method has reasonable sensitivity and flexibility to be applied as a detection and quantification method before the discovery of variants with more mutated VP1 gene.

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Section: Introductionmentioning

confidence: 99%

Updating Sacbrood Virus Quantification PCR Method Using a TaqMan-MGB Probe

Huang

Zhang

Mehmood

et al. 2021

Veterinary Sciences

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Colorimetric biosensors for point-of-care virus detections

Zhao

Wong

Zheng

et al. 2020

Materials Science for Energy Technologies

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a b s t r a c tColorimetric biosensors can be used to detect a particular analyte through color changes easily by naked eyes or simple portable optical detectors for quantitative measurement. Thus, it is highly attractive for point-of-care detections of harmful viruses to prevent potential pandemic outbreak, as antiviral medication must be administered in a timely fashion. This review paper summaries existing and emerging techniques that can be employed to detect viruses through colorimetric assay design with detailed discussion of their sensing principles, performances as well as pros and cons, with an aim to provide guideline on the selection of suitable colorimetric biosensors for detecting different species of viruses. Among the colorimetric methods for virus detections, loop-mediated isothermal amplification (LAMP) method is more favourable for its faster detection, high efficiency, cheaper cost, and more reliable with high reproducible assay results. Nanoparticle-based colorimetric biosensors, on the other hand, are most suitable to be fabricated into lateral flow or lab-on-a-chip devices, and can be coupled with LAMP or portable PCR systems for highly sensitive on-site detection of viruses, which is very critical for early diagnosis of virus infections and to prevent outbreak in a swift and controlled manner.

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Current Status of Loop-Mediated Isothermal Amplification Technologies for the Detection of Honey Bee Pathogens

2021

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Approximately one-third of the typical human Western diet depends upon pollination for production, and honey bees (Apis mellifera) are the primary pollinators of numerous food crops, including fruits, nuts, vegetables, and oilseeds. Regional large scale losses of managed honey bee populations have increased significantly during the last decade. In particular, asymptomatic infection of honey bees with viruses and bacterial pathogens are quite common, and co-pathogenic interaction with other pathogens have led to more severe and frequent colony losses. Other multiple environmental stress factors, including agrochemical exposure, lack of quality forage, and reduced habitat, have all contributed to the considerable negative impact upon bee health. The ability to accurately diagnose diseases early could likely lead to better management and treatment strategies. While many molecular diagnostic tests such as real-time PCR and MALDI-TOF mass spectrometry have been developed to detect honey bee pathogens, they are not field-deployable and thus cannot support local apiary husbandry decision-making for disease control. Here we review the field-deployable technology termed loop-mediated isothermal amplification (LAMP) and its application to diagnose honey bee infections.

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Rapid detection of sacbrood virus (SBV) by one-step reverse transcription loop-mediated isothermal amplification assay

Cited by 5 publications

References 4 publications

Updating Sacbrood Virus Quantification PCR Method Using a TaqMan-MGB Probe

Updating Sacbrood Virus Quantification PCR Method Using a TaqMan-MGB Probe

Colorimetric biosensors for point-of-care virus detections

Current Status of Loop-Mediated Isothermal Amplification Technologies for the Detection of Honey Bee Pathogens

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