Non-polio Enterovirus (NPEV) and Human Parechovirus (HPeV) Molecular Detection and Typing

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Requisition Forms

Reference Details

Description:

Molecular Detection and Genotyping by RT-semi-nested PCR and sequencing (if poliovirus is suspected refer to the Guide to Services for Polioviruses).

Test Category:
Molecular Detection and Genotyping
Pathogen:
Parechoviruses
Illnesses and Diseases:
  • Acute flaccid paralysis
  • Acute haemorrhagic conjunctivitis
  • Aseptic meningitis/encephalitis
  • Gastrointestinal illnesses
  • Hand foot and mouth disease
  • Myocarditis
  • Non-specific febrile illness with or without rash
  • Pleurodynia
  • Sepsis-like picture in neonates
  • Upper respiratory tract infections
Specimen:
  • Viral Isolates/Culture -≥ 0.5 mL of infected cells and media.
  • CSF - ≥ 0.2 mL.
  • Serum/Plasma - Not recommended.
  • Frozen Whole Blood - Not recommended-this sample type will be rejected for testing.
  • Respiratory NP/OP swab, wash or aspirate - ≥ 0.5 mL.
  • Stool -  ≥ 1.0 g (solid) or ≥ 0.5 mL (liquid).  Virus present for longer than other specimen types.
  • Rectal swab
Collection Method:
  • Viral Isolates/Culture - Collect in sterile leak proof container.
  • CSF - Collect in sterile leak proof container, no special medium required.
  • Respiratory NP/OP swab, wash or aspirate - For washes and aspirates collect in sterile leak proof container, no special medium required. For swabs, store in viral transport medium.
  • Stool -  Collect in sterile leak proof container, no special medium required.
  • Rectal swab - Store in viral transport medium.
Specimen Processing, Storage and Shipping:

Store samples frozen at ≤-20°C until shipped for testing. Ship frozen on dry ice. Package Enterovirus/Parechovirus samples in a separate shipping container from other samples being sent to the NML.

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 enterovirus or parechovirus infections.

Accompanying Documentation:

Completed Enteroviruses and Enteric viruses requisition form. If possible include clinical and travel history and any enterovirus or parechovirus lab test results completed at local or provincial laboratories.

Comments:

Labs that perform their own enterovirus and/or parechovirus PCR testing should submit positive samples for genotyping surveillance.

Methods and Interpretation of Results:

Laboratory surveillance for NPEVs and HPeV is based on published protocols that take advantage of the correlation between the partial VP1 sequence and the enterovirus (EV) serotype and parechovirus genotype. Partial EV and HPeV VP1 sequences are amplified from clinical isolates or direct specimens by RT- semi nested PCRs (RT-snPCR) using generic primers that target all human enterovirus serotypes and parechovirus genotypes to date. The amplicons generated from these RT-snPCRs are sequenced and the virus identity is determined by entering the sequence into a web-based automated genotyping tool for enteroviruses and noroviruses. A negative result does not rule out the presence of enterovirus. A negative result may be due to PCR inhibitors present in the patient sample or the viral load in the sample is below the level of detection of the assay.

Turnaround Time:

21 calendar days. Stat testing available upon request.

Contact:
Phone: (204) 789-2022
Fax: (204) 789-2082
Additional Contacts
Elsie Grudeski
Phone: 204-789-6067
Fax: 204-789-2082
References:
  1. W.A. Nix, M. S. Oberste, and M.A. Pallansch. Sensitive, Semi-nested PCR Amplification of VP1 Sequences for Direct Identification of All Enterovirus Serotypes from Original Clinical Specimens. 2006 J. Clin. Microbiol. 44: 2698-2704.
  2. W.A. Nix, K. Maher, M.A. Pallansch, M.S. Oberste. Parechovirus typing in clinical specimens by nested or semi-nested PCR coupled with sequencing. 2010 J. Clin. Virol. 48: 202-207.
  3. A. Kroneman A, H. Vennema, K. Deforche, H.v.d. Avoort, S. Peñaranda, M.S, Oberste, J. Vinjé, M. Koopmans. An automated genotyping tool for enteroviruses and noroviruses. 2011 J Clin Virol. Jun; 51(2):121-5).
Guidelines:
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