Genotyping by RT-PCR

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Accredited by the Standards Council of Canada to Laboratory no. 594 - CAN-P-4E (ISO/IEC 17025)

Requisition Forms

Reference Details

Description:

Genotyping of Influenza A virus (H1 and H3), Influenza B virus, and Avian Influenza A virus (H1 to H16) by real-time RT-PCR and/or RT-PCR and sequencing.

Test Category:
Genotyping
Pathogen:
Influenza virus
Illnesses and Diseases:
  • Influenza
Specimen:

Respiratory specimens (nasopharyngeal swab, throat swab, etc.) or viral isolates. Volume required: 1-2 mL.

Collection Method:

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

Specimen Processing, Storage and Shipping:

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

Transportation of Dangerous Goods:

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.

For additional guidance on the transport of infectious substances in other languages, please click on the link below.

http://www.who.int/ihr/capacity-strengthening/infectious-substances/en/

Patient Criteria:

Suspected or confirmed influenza virus infection.

Accompanying Documentation:

Completed requisition (current WHO Influenza Program – Antigenic Subtyping 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: passage history, patient travel history, extent of influenza activity, and date of collection relative to antiviral use.

Comments:

Authorization must be obtained for genotyping by RT-PCR and sequencing on Influenza A virus (H1 and H3), Influenza B virus, Avian Influenza A virus (H1 to H16).  Please contact the Influenza and Respiratory Viruses laboratory before sending specimens.  Samples received three months after collection date will not be tested unless prior arrangements have been made with the Section Chief.

Methods and Interpretation of Results:

Nucleic acids are extracted from submitted specimens/isolates. Reverse Transcriptase Polymerase Chain Reaction (RT-PCR) is performed on extracted nucleic acids with real-time detection or visual confirmation (gel electrophoresis) of PCR products. Conventional PCR products may be sequenced and aligned with reference sequences to determine the genotype.

Turnaround Time:

14 calendar days.

Contact:
Phone: (204) 789-6045
Fax: (204) 789-2082
References:
  1. Real-Time RT-PCR (rRT-PCR) Protocol for Influenza. CDC. Revised, 2007.
  2. Fouchier R. et al. Detection of Influenza A Viruses from Different Species by PCR Amplification of Conserved Sequences in the Matrix Gene. J. Clin. Microbiol. (2000) 38: 4096-4101