About: Development of a loop-mediated isothermal amplification assay for the detection and quantification of epizootic epitheliotropic disease virus (salmonid herpesvirus-3)

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About: Development of a loop-mediated isothermal amplification assay for the detection and quantification of epizootic epitheliotropic disease virus (salmonid herpesvirus-3) Goto Sponge NotDistinct Permalink

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type	Academic Article research paper schema:ScholarlyArticle
isDefinedBy	Covid-on-the-Web dataset
has title	Development of a loop-mediated isothermal amplification assay for the detection and quantification of epizootic epitheliotropic disease virus (salmonid herpesvirus-3)
Creator	Faisal, Mohamed Standish, Isaac Zhang, Qingli Glenney, Gavin Loch, Thomas Shavalier, Megan
Source	Elsevier; Medline; PMC
abstract	Epizootic Epitheliotropic Disease Virus (EEDV; Salmonid Herpesvirus-3) causes a serious disease hatchery-reared lake trout (Salvelinus namaycush), threatening restoration efforts of this species in North America. The current inability to replicate EEDV in vitro necessitates the search for a reproducible, sensitive, and specific assay that allows for its detection and quantitation in a time- and cost-effective manner. Herein, we describe a loop-mediated isothermal amplification (LAMP) assay that was developed for the quantitative detection of EEDV in infected fish tissues. The newly developed LAMP reaction was optimized in the presence of calcein, and the best results were produced using 2 mM MgCl(2), 1.8 mM dNTPs and at an incubation temperature of 67.1 °C. This method was highly specific to EEDV, as it showed no cross-reactivity with several fish viruses, including Salmonid Herpesvirus(-1), -2, -4, and -5, Infectious Pancreatic Necrosis Virus, Spring Viremia of Carp Virus, Infectious Hematopoietic Necrosis Virus, Golden Shiner Reovirus, Fathead Minnow Nidovirus, and Viral Hemorrhagic Septicemia Virus. The analytical sensitivity of the EEDV-LAMP method was estimated to be as low as 16 copies of plasmid per reaction. When infected fish tissue was used, a positive reaction could be obtained when an infected gill tissue sample that contained 430 viral copies/μg was diluted up to five orders of magnitude. The sensitivity and specificity of the newly developed LAMP assay compared to the SYBR Green qPCR assay were 84.3% and 93.3%, respectively. The quantitative LAMP for EEDV had a correlation coefficient (R(2) = 0.980), and did not differ significantly from the SYBR Green quantitative PCR assay (p > 0.05). Given its cost- and time-effectiveness, this quantitative LAMP assay is suitable for screening lake trout populations and for the initial diagnosis of clinical cases.
has issue date	2018-11-13(xsd:dateTime)
bibo:doi	10.1016/j.jviromet.2018.11.006
bibo:pmid	30444983
has license	no-cc
sha1sum (hex)	805440d5e0c269e5c488937c6636ab0a2446b979
schema:url	https://doi.org/10.1016/j.jviromet.2018.11.006
resource representing a document's title	Development of a loop-mediated isothermal amplification assay for the detection and quantification of epizootic epitheliotropic disease virus (salmonid herpesvirus-3)
has PubMed Central identifier	PMC7119762
has PubMed identifier	30444983
schema:publication	J Virol Methods
resource representing a document's body	covid:805440d5e0c269e5c488937c6636ab0a2446b979#body_text
is schema:about of	named entity 'developed' named entity 'The' named entity 'Spring' named entity 'Disease' named entity 'specific' named entity 'LAMP' named entity 'reaction' named entity 'ORDERS' named entity 'BEST' named entity 'RESULTS' named entity 'COMPARED' named entity 'NORTH AMERICA' named entity 'CALCEIN' named entity 'CURRENT' named entity 'ANALYTICAL' named entity 'DISEASE VIRUS' named entity 'EFFECTIVENESS' named entity 'POSITIVE' named entity 'ASSAY' named entity 'CORRELATION COEFFICIENT' named entity 'VIRAL' named entity '2 MM' named entity 'FISH TISSUE' named entity 'SPRING VIREMIA OF CARP VIRUS' named entity 'VIRAL HEMORRHAGIC SEPTICEMIA VIRUS' named entity 'NEWLY' named entity 'SENSITIVE' named entity 'INFECTIOUS PANCREATIC NECROSIS VIRUS' named entity 'GOLDEN SHINER REOVIRUS' named entity 'HATCHERY' named entity 'SENSITIVITY AND SPECIFICITY' named entity 'DISEASE' named entity '1.8' named entity '0.05' named entity 'SENSITIVITY' named entity 'INFECTED' named entity 'EPIZOOTIC' named entity 'QUANTIFICATION' named entity 'DISEASE VIRUS' named entity 'HERPESVIRUS' named entity 'DETECTION' named entity 'LOOP-MEDIATED ISOTHERMAL AMPLIFICATION' named entity 'DEVELOPMENT' named entity 'DID' named entity 'COPIES' named entity 'SPECIFIC' named entity 'DETECTION' named entity 'SCREENING' named entity 'INABILITY' named entity 'LAMP' named entity 'REACTION' named entity 'EFFECTIVE' named entity 'VIRUSES' named entity 'SUITABLE' named entity 'USED' named entity 'ISOTHERMAL AMPLIFICATION ASSAY' named entity 'EPIZOOTIC' named entity 'METHOD' named entity 'IN VITRO' named entity 'HIGHLY' named entity 'REACTIVITY' named entity 'CROSS' named entity '430' named entity 'SYBR GREEN' named entity 'CAUSES' named entity 'GILL' named entity 'COULD BE' named entity 'INFECTIOUS HEMATOPOIETIC NECROSIS VIRUS' named entity 'TISSUE SAMPLE' named entity 'LAMP METHOD' named entity 'BCG' named entity 'FISH' named entity 'FATHEAD MINNOW' named entity 'RESTORATION' named entity 'TIME' named entity 'QPCR ASSAY'

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