Molecular Detection and Genotyping

Molecular detection of the mumps virus by real-time RT-PCR and genotyping of positive specimens.

Urine (collected within 2 weeks post onset), parotid gland/buccal swabs or throat swabs, saliva (collected within 5 days post onset). Note: specimens collected later will still be accepted however the assay sensitivity will not be optimal. CSF may be submitted for meningitis/encephalitis cases. Viral isolates may also be sent.

Collect 50 mL (minimum 10 mL) of urine in sterile container. For swabs, collect specimens using sterile swabs approved for virus isolation and place in 2 -3 mL of viral transport medium (VTM) for a minimum of 1 hour. For saliva, mix with equal part VTM. For viral isolate, collect infected cells and media and submit 1.0 mL at minimum. For CSF, submit at minimum 0.5 mL in a sterile collection vial.

Process urine by centrifuging at 2500 x g for 15 minutes at 4EC. Resuspend the sediment in 2.0 mL VTM. For swab specimens, remove the swab. For all specimens, store at 4°C and ship on wet ice for arrival at the NML within 2 days from collection. Otherwise, freeze at -70°C and ship frozen on dry ice. Viral Isolates should be shipped frozen (-70°C) 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.

High fever, fatigue and unilteral or bilateral parotitis. Suspect mumps meningitis/encephalitis cases.

Completed Measles, Mumps, and Rubella requisition form. Include date of symptom onset, date of last mumps vaccination (if recent) and travel history.

Labs that perform their own mumps RT-PCR testing should submit positive samples for genotype surveillance. It is important to provide a complete patient history (including travel) for accurate genotype surveillance.

Extracted RNA from samples is tested by real-time RT-PCR for the mumps Fusion gene (2). A positive RT-PCR result is laboratory confirmation of mumps infection. The WHO standardised region, the small hydrophobic (SH) gene, of positive samples is amplified by conventional RT-PCR and sequenced to determine the genotype of the mumps virus (1).

Molecular detection: 7 calendar days. Genotyping: 21 calendar days. Stat testing is available upon request.

1. World Health Organization. Mumps virus nomenclature update: 2012. Weekly Epidemiological Record 2012; 87: 217-224.
2. Uchida K, M Shinohara, S Shimada et al. Rapid and sensitive detection of mumps virus RNA directly from clinical samples by real-time PCR. J Med Virology 2005;75:470–4.
3. Public Health Agency of Canada. Laboratory guidelines for the diagnosis of mumps. CCDR. 2010; 36S1: 37-41.
4. Tipples G, J Hiebert. Detection of measles, mumps and rubella viruses. Methods Mol Biol. 2011; 665: 183-193.