Document Detail


A Simple-Probe real-time PCR assay for genotyping reassorted and non-reassorted isolates of Crimean-Congo hemorrhagic fever virus in southern Africa.
MedLine Citation:
PMID:  20599559     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Crimean-Congo hemorrhagic fever (CCHF) is a tick-borne viral zoonosis distributed widely in Africa, Asia, Russia and the Balkans. Occurrence of segment reassortment has been established and may be associated with pathogenicity. The ability to distinguish between an infection with a reassortant and a non-reassortant variant may have prognostic value. In this study the use of Simple-Probe technology and real-time PCR was investigated to detect rapidly CCHF viral nucleic acid and to distinguish between reassorted and non-reassorted variants of southern African CCHF virus (CCHFV) isolates. A Simple-Probe was designed based on multiple alignments of 19 CCHFV partial M segment sequences that included reassorted and non-reassorted isolates from two phylogenies. Real-time PCR followed by probe-specific melting-curve analysis allowed differentiation of three selected isolates representative of two phylogenetically distinct groups, SPU 431/85 (group IV), 383/87 (group IV) and 103/87 (group III) based on melting temperatures. Within group IV there are two distinct lineages which were represented in the optimization. A further 16 clinical isolates could be genotyped into groups III and IV using the Simple-Probe real-time PCR assay developed in this study. This assay provides a reliable and sensitive tool for the differentiation between reassorted and non-reassorted variants of CCHFV which finds application for diagnosis and epidemiologic and surveillance studies.
Authors:
Kulsum Kondiah; Robert Swanepoel; Janusz T Paweska; Felicity J Burt
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Publication Detail:
Type:  Evaluation Studies; Journal Article; Research Support, Non-U.S. Gov't     Date:  2010-06-25
Journal Detail:
Title:  Journal of virological methods     Volume:  169     ISSN:  1879-0984     ISO Abbreviation:  J. Virol. Methods     Publication Date:  2010 Oct 
Date Detail:
Created Date:  2010-08-30     Completed Date:  2010-12-08     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  8005839     Medline TA:  J Virol Methods     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  34-8     Citation Subset:  IM    
Copyright Information:
Copyright (c) 2010 Elsevier B.V. All rights reserved.
Affiliation:
Department of Biotechnology, University of Johannesburg, PO Box 17011, Doornfontein 2028, South Africa.
Data Bank Information
Bank Name/Acc. No.:
GENBANK/FJ435387;  FJ435388;  FJ435389;  FJ435390;  FJ435391;  FJ435392;  FJ435393;  FJ435394;  FJ435395;  FJ435396;  FJ435397;  FJ435398;  FJ435399;  FJ435400;  FJ435401;  FJ435402;  FJ435403;  FJ435404;  FJ435405;  FJ435406;  FJ435407;  FJ435408;  FJ435409
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MeSH Terms
Descriptor/Qualifier:
Africa, Southern
Animals
Genotype
Hemorrhagic Fever Virus, Crimean-Congo / classification*,  genetics*,  isolation & purification
Hemorrhagic Fever, Crimean / virology*
Humans
Molecular Sequence Data
Polymerase Chain Reaction / methods*
RNA, Viral / genetics
Reassortant Viruses / classification,  genetics,  isolation & purification
Sensitivity and Specificity
Sequence Analysis, DNA
Transition Temperature
Virology / methods*
Chemical
Reg. No./Substance:
0/RNA, Viral

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


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