NDLI: Development of multiplexed real-time quantitative polymerase chain reaction assay for detecting human adenoviruses.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Huang, Meei-Li Nguy, Long Ferrenberg, James Boeckh, Michael Cent, Anne Corey, Lawrence
Description	Country affiliation: United States Author Affiliation: Huang ML ( Department of Laboratory Medicine, University of Washington, Seattle, WA 98195, USA.)
Abstract	Adenoviruses (AdVs) have been associated with a wide variety of human disease and are increasingly recognized as viral pathogens that can cause significant morbidity and mortality in immunocompromised patients. Early detection of AdV DNA in plasma and sterile fluids has been shown to be useful for identifying patients at risk for invasive AdV disease. Because of the large number of existing Adv types, few real-time quantitative AdV polymerase chain reaction (PCR) assays published effectively cover all AdV types. We designed a series of AdV PCR primers and probes and empirically multiplexed them into 2 separate real-time PCR assays to quantitatively detect all 49 serotypes of human AdV (types 1-49) available from American Type Culture Collection. We then subsequently multiplexed all the primers and probes into 1 reaction. The sensitivity of these assays was determined to be less than 10 copies per reaction (500 copies/mL plasma). In a retrospective evaluation, we detected all 84 clinical AdV isolates isolated in cell culture from patients undergoing hematopoietic stem cell transplantation between 1981 and 1987. Prospective analysis of 46 consecutive clinical samples submitted for AdV testing showed greater sensitivity and equal specificity of the AdV PCR than viral culture. This real-time PCR assay allows rapid, sensitive, and specific quantification of all currently defined AdVs into either 2 or 1 multiplex assay for clinical samples.
File Format	HTM / HTML
ISSN	07328893
e-ISSN	18790070
DOI	10.1016/j.diagmicrobio.2008.06.009
Journal	Diagnostic Microbiology and Infectious Disease
Issue Number	3
Volume Number	62
Language	English
Publisher	Elsevier
Publisher Date	2008-11-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Discipline Microbiology Discipline Communicable Diseases Adenovirus Infections, Human Virology Adenoviruses, Human Isolation & Purification Polymerase Chain Reaction Diagnosis Genetics Dna Primers Dna Probes Dna, Viral Molecular Diagnostic Techniques Prospective Studies Retrospective Studies Sensitivity And Specificity Serotyping Evaluation Studies Research Support, N.i.h., Extramural
Content Type	Text
Resource Type	Article
Subject	Infectious Diseases Microbiology (medical)

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1	Ministry of Education (GoI), Department of Higher Education	Sanctioning Authority	https://www.education.gov.in/ict-initiatives
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4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
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