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Development and application of a one-step real-time RT-PCR using a minor-groove-binding probe for the detection of a novel bunyavirus in clinical specimens.
MedLine Citation:
PMID:  23212930     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
A highly sensitive one-step real-time RT-PCR method using a minor-groove-binding (MGB) probe was developed for detection and quantitation of severe febrile with thrombocytopenia syndrome virus (SFTSV). The assay could discriminate SFTSV infection from other related viral diseases in human with a minimum detection limit of 10 viral RNA copies/µl and was 1,000 times more sensitive than the conventional PCR. Strong linear correlations (r(2)  > 0.99) between the C(t) values and viral RNA standards over a linear range were obtained. The coefficients of variation of intra- and inter-assay reproducibility were both less than 2%. The RT-PCR was also shown to be highly specific, as no positive signals were detected for other related viruses. Evaluation of this assay with serum samples from laboratory confirmed cases and healthy donors showed 100% clinical diagnostic sensitivity and over 99% specificity. Clinical application with samples from 287 patients admitted to the hospital with suspected SFTSV infection showed that 15% were infected by SFTSV. This assay was rapid, requiring just over 2 hr, including the nucleic acid extraction step. J. Med. Virol. © 2012 Wiley Periodicals, Inc.
Authors:
Zhifeng Li; Lunbiao Cui; Minghao Zhou; Xian Qi; Changjun Bao; Jianli Hu; Jun Shan; Bin Wu; Shenjiao Wang; Xiling Guo; Yongjun Jiao; Fenyang Tang; Hua Wang
Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2012-12-4
Journal Detail:
Title:  Journal of medical virology     Volume:  -     ISSN:  1096-9071     ISO Abbreviation:  J. Med. Virol.     Publication Date:  2012 Dec 
Date Detail:
Created Date:  2012-12-5     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  7705876     Medline TA:  J Med Virol     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Copyright Information:
Copyright © 2012 Wiley Periodicals, Inc.
Affiliation:
Department of Acute Infectious Diseases Control and Prevention, Jiangsu Provincial Center for Disease Control and Prevention, Nanjing, China.
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