Evaluation of type-specific real-time PCR assays using the LightCycler and J.B.A.I.D.S. for detection of adenoviruses in species HAdV-C

Sporadically, HAdVs from species HAdV-C are detected in acute respiratory disease outbreaks. To rapidly type these viruses, we designed real-time PCR assays that detect and discriminate between adenovirus types HAdV-C1, -C2, -C5, and -C6. Sixteen clinical isolates from the California Department of P...

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Bibliographic Details
Published in:	PloS one 2011-10, Vol.6 (10), p.e26862-e26862
Main Authors:	Jones, Morris S, Hudson, Nolan Ryan, Gibbins, Carl, Fischer, Stephen L
Format:	Article
Language:	eng
Subjects:	Adenoviridae - genetics Adenoviridae - isolation & purification Adenovirus Infections, Human - diagnosis Adenoviruses Adenoviruses, Human - genetics Adenoviruses, Human - isolation & purification Analysis Assaying Bacteria Biology Clinical isolates Deoxyribonucleic acid Diagnostic software Diagnostic systems Discrimination DNA Epidemics Genomes Health aspects Humans Identification Infectious diseases Influenza Laboratories Limit of Detection Medicine Molecular Sequence Data Outbreaks Pneumonia Public health Real time Real-Time Polymerase Chain Reaction - instrumentation Real-Time Polymerase Chain Reaction - methods Real-Time Polymerase Chain Reaction - standards Regression analysis Respiratory diseases Respiratory syncytial virus Respiratory tract diseases Sensitivity and Specificity Species Training Viral proteins Virology Viruses
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Summary:	Sporadically, HAdVs from species HAdV-C are detected in acute respiratory disease outbreaks. To rapidly type these viruses, we designed real-time PCR assays that detect and discriminate between adenovirus types HAdV-C1, -C2, -C5, and -C6. Sixteen clinical isolates from the California Department of Public Health were used to validate the new assays. Type-specific TaqMan real-time PCR assays were designed and used independently to successfully identify 16 representative specimens. The lower limit of detection for our LightCycler singleplex real-time PCR assays were calculated to be 100, 100, 100, and 50 genomic copies per reaction for HAdV-C1, HAdV-C2, HAdV-C5 and HAdV-C6, respectively. The results for the singleplex J.B.A.I.D.S. assays were similar. Our assays did not cross-react with other adenoviruses outside of species HAdV-C, respiratory syncytial virus, influenza, or respiratory disease causing bacteria. These assays have the potential to be useful as diagnostic tools for species HAdV-C infection.
ISSN:	1932-6203 1932-6203