title: 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.
abstract: 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.
authors: Viral and Rickettsial Disease Laboratory, California Department of Health Services, Richmond, California, United States of America. morris.jones@cdph.ca.gov
authors: Morris S
authors: MS
authors: Jones
authors: Nolan Ryan
authors: NR
authors: Hudson
authors: Carl
authors: C
authors: Gibbins
authors: Stephen L
authors: SL
authors: Fischer
mesh_terms: D000256:Adenoviridae / Q000235:genetics / Q000302:isolation & purification*; D000258:Adenovirus Infections, Human / Q000175:diagnosis; D000260:Adenoviruses, Human / Q000235:genetics / Q000302:isolation & purification; D006801:Humans; D057230:Limit of Detection; D008969:Molecular Sequence Data; D060888:Real-Time Polymerase Chain Reaction / Q000295:instrumentation* / Q000379:methods* / Q000592:standards; D012680:Sensitivity and Specificity