Document Detail


Loop mediated isothermal amplification combined with nucleic acid lateral flow strip for diagnosis of cyprinid herpes virus-3.
MedLine Citation:
PMID:  19781627     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
An improved loop mediated isothermal amplification (LAMP) assay for rapid, sensitive and specific detection of cyprinid herpes virus-3 (CyHV-3), also known as koi herpes virus (KHV), was developed. The lower detection limit of the CyHV-3-LAMP assay is 10 fg DNA which equivalent to 30 copies of CyHV-3 genome. Nucleic acid lateral flow assay was used for visual detection of the LAMP products. The LAMP- nucleic acid lateral flow assay relies on DNA hybridization technology and antigen-antibody reactions in combination with LAMP. For application of this assay, the biotinylated LAMP product was hybridized with a FITC-labelled specific probe for 5 min. The resulting DNA complex could be visualised as purple band at the strip test line within 5 min of sample exposure. The nucleic acid lateral flow analysis of the LAMP product was equivalent in sensitivity but more rapid than the conventional agarose gel electrophoresis. The combination of LAMP assay with the nucleic acid lateral flow analysis can simplify the diagnosis and screening of CyHV-3 as it is simple, requires very little training, does not require specialized equipment such as a thermal cycler, the results are read visually with no need to run a gel and has a high sensitivity and specificity.
Authors:
Hatem Soliman; Mansour El-Matbouli
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Publication Detail:
Type:  Journal Article     Date:  2009-09-23
Journal Detail:
Title:  Molecular and cellular probes     Volume:  24     ISSN:  1096-1194     ISO Abbreviation:  Mol. Cell. Probes     Publication Date:  2010 Feb 
Date Detail:
Created Date:  2010-02-03     Completed Date:  2010-04-22     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  8709751     Medline TA:  Mol Cell Probes     Country:  England    
Other Details:
Languages:  eng     Pagination:  38-43     Citation Subset:  IM    
Copyright Information:
Copyright 2009 Elsevier Ltd. All rights reserved.
Affiliation:
Fish Medicine and Livestock Management, University of Veterinary Medicine, Vienna, Austria. hatemtoughan@hotmail.com
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MeSH Terms
Descriptor/Qualifier:
Animals
Cell Line, Tumor
Cyprinidae / virology*
Herpesviridae / genetics,  isolation & purification*
Herpesviridae Infections / diagnosis,  virology
Humans
Nucleic Acid Amplification Techniques*
Polymerase Chain Reaction

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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