Genotyping by PCR and Sequencing

Genotyping of Adenovirus spp. by PCR and sequencing.

Respiratory specimens (nasopharygeal swab, throat swab, other), or viral isolate (1.0-2.0 mL). Other patient specimens (eg. tissues) may be tested, although the test works best with respiratory specimens.

For respiratory specimens, use sterile swabs and swab nasal cavity, throat, etc. Place swabs in 2-3 mL viral transport medium.

Keep samples refrigerated until shipped for testing. Ship on dry ice.

Shipping of specimens shall be done by a TDG certified individual in accordance with TDG regulations. For additional information regarding classification of specimens for the purposes of shipping, consult either Part 2 Appendix 3 of the TDG Regulations or section 3.6.2 of the IATA Dangerous Goods Regulations as applicable.

Suspected or confirmed Adenovirus infection.

Completed requisition (current Adenovirus Genotyping requisition form). Sender lab name, address, and telephone number must be provided for samples to be processed. Also include identifier (sender lab number), patient date of birth, patient gender, type of specimen, and date collected. Under additional information, please include if possible/applicable patient travel history and date of collection relative to antiviral use.

Authorization must be obtained for genotyping by PCR and sequencing for Adenovirus spp. Please contact the Influenza and Respiratory Viruses laboratory before sending specimens.

Nucleic acids are extracted from submitted specimens/isolates. Polymerase Chain Reaction (PCR) with Adenovirus-specific primers is performed on extracted nucleic acids. If initial PCR is unsuccessful, nested PCR with additional Adenovirus-specific primers is performed. PCR products are sequenced and aligned with reference sequences to determine the genotype.

14 calendar days.

1. X Lu, DD Erdman. Molecular typing of human adenoviruses by PCR and sequencing of a partial region of the hexon gene. Arch Virol (2006) 151: 1587-1602.