Specimen Collection, Storage, and Shipment

Collecting Specimens for Suspected Mumps Cases

If it has been <3 days since symptom onset, collect a buccal swab specimen for detection of viral RNA by RT-qPCR. If it has been >3 days since symptom onset, collect a buccal swab specimen for RT- qPCR and a serum specimen for IgM detection. If the patient has orchitis/oophoritis, mastitis, pancreatitis, hearing loss, meningitis, or encephalitis, collect a buccal specimen for RT-qPCR, a urine specimen for RT-qPCR, and a serum specimen for IgM detection, regardless of days since symptom onset.

At the onset of a suspected mumps outbreak, patients suspected to have mumps should be tested by RT-qPCR to confirm mumps and rule out other possible etiologies. However, once a mumps outbreak is confirmed, jurisdictions should consider alternate strategies to ensure efficient use of resources available for laboratory testing.

The CSTE clinical case definition for mumps designates cases with a positive RT-PCR result as confirmed, while cases that are IgM positive are classified as probable.

Shipping Specimens for Suspected Mumps Cases

Please contact your state health department to determine where to submit specimens and how to ship them.

If instructed to send specimens to CDC from within the U.S., you must use the CDC Specimen Submission Form (50.34). Please provide a submission form for each specimen submitted.

Please be sure to include the following information:

  • Contact name with telephone number and email address
  • Case ID and specimen ID from the submitting laboratory associated with the specimen
  • Type of specimen collected
  • Date specimen collected
  • MMR vaccination history [date(s) if known]
  • Clinical signs and symptoms
  • Patient’s date of birth or age
  • Onset date of parotitis or jaw swelling if applicable
  • Any previous test results (if known)
Ship to:
Centers for Disease Control and Prevention
STAT Unit #81
Attn: Dr. Paul Rota
1600 Clifton Road NE
Atlanta, GA 30329
Contact Paul Rota by telephone at 404-639-4181 or by e-mail at prota@cdc.gov for additional information and to provide shipment details.

Oral or buccal swab samples

Video: “Collecting a Buccal Swab Clinical Specimen for Mumps Diagnostic Testing.”
video: Collecting a Buccal Swab Clinical Specimen for Mumps Diagnostic Testing.

This short video demonstrates how to correctly collect and transport a buccal swab (the preferred sample) for the detection of mumps virus RNA.

Collect oral or buccal swab samples as soon as mumps disease is suspected. RT-PCR has the greatest diagnostic sensitivity when samples collected with 3 days of symptom onset.

The buccal or oral swab specimens are obtained by massaging the parotid gland area for 30 seconds prior to swabbing the area around Stensen’s duct. A commercial product designed for the collection of throat specimens or a flocked polyester fiber swab can be used. Synthetic swabs are preferred over cotton swabs, which may contain substances that are inhibitory to enzymes used in RT-PCR. Flocked synthetic swabs appear to be more absorbent and elute samples more efficiently.

Processing the swabs within 24 hours of collection will enhance the sensitivity of both the RT-PCR and virus isolation techniques.

Swabs should be placed in 2 ml of standard viral transport medium (VTM)[1]. Allow the swab to remain in VTM for at least 1 hour (4°C). Ream the swab around the rim of the tube to retain cells and fluid in the tube. The swab can be broken off and left in the tube or discarded.

Storage and shipment: Following collection, samples should be maintained at 4°C and shipped on cold packs (4°C) within 24 hours. If there is a delay in shipment, the sample is best preserved by freezing at −70°C. Frozen samples should be shipped on dry ice.

Urine specimens

Urine is not the optimal specimen for RT-qPCR and is not preferred as a specimen for patients with parotitis, even for those patients presenting more than 3 days since symptom onset (Jin et al, 2007, Tan et al, 2011, Maillet et al, 2015, L’huillier et al, 2018, Krause et al, 2006, Hatchette et al, 2009, Nunn et al, 2018). Testing urine during an outbreak does not significantly increase diagnostic yield and can negatively impact testing resources.

However, in patients presenting with mumps complications (orchitis, oophoritis, nephritis, encephalitis, meningitis) collecting a urine specimen in addition to an oral specimen might help to confirm mumps infection.

Obtaining urine in patients with complications may lead to a better understanding of the utility of urine as a specimen for RT-qPCR in these cases.

A minimum volume of 50 ml of urine should be collected in a sterile container and then processed by centrifuging at 2500 × g for 15 minutes at 4°C. The sediment should be resuspended in 2 ml of VTM.

Storage and Shipment: Store urine sediment in VTM at 4°C and, if possible, ship on cold packs within 24 hours. The best method for preserving mumps virus in processed (centrifuged) urine is to freeze the sample at −70°C and ship on dry ice. See information on shipping, including where to send samples for testing.

Serology (serum samples)

If it has been >3 days after symptom onset, collect 7–10 ml of blood in a red-top or serum-separator tube (SST) for IgM detection. Centrifuge blood collection tubes (10 min at 2200 – 2500 rpm) to separate serum from clot. Gel separation tubes should be centrifuged no later than 2 hours after collection. Aseptically transfer serum to a sterile tube that has an externally threaded cap with an o–ring seal. Store specimens at 4°C and ship on wet ice packs. Do not freeze tubes containing whole blood.  Hemolyzed and lipemic serum and plasma are noted and tested, usually without apparent interferences.

If the serum sample collected >3 days after parotitis onset is IgM negative, and the case has a negative (or not done) result for RT-qPCR, and there is a strong suspicion of mumps a second serum sample collected 5–10 days after symptom onset is recommended because, in some cases, the IgM response is not detectable until 5 days after symptom onset.

A single serum specimen tested for mumps specific IgG is not useful for diagnosing acute mumps infections. A fourfold rise or seroconversion is rarely demonstrated between paired serum samples for mumps since IgG is typically present at symptom onset (see serology FAQ).

References

  • Hatchette TF, Mahony JB, Chong S, LeBlanc JJ. Difficulty with mumps diagnosis: what is the contribution of mumps mimickers? J Clin Virol 2009; 46:381-3.
  • Jin L, Feng Y, Parry R, Cui A, Lu Y. Real-time PCR and its application to mumps rapid diagnosis. J Med Virol 2007; 79:1761-7.
  • Krause CH, Eastick K, Ogilvie MM. Real-time PCR for mumps diagnosis on clinical specimens–comparison with results of conventional methods of virus detection and nested PCR. J Clin Virol 2006; 37:184-9.
  • L’Huillier AG, Eshaghi A, Racey CS, et al. Laboratory testing and phylogenetic analysis during a mumps outbreak in Ontario, Canada. Virol J 2018; 15:98.
  • Maillet M, Bouvat E, Robert N, et al. Mumps outbreak and laboratory diagnosis. J Clin Virol 2015; 62:14-9.
  • Nunn A, Masud S, Krajden M, Naus M, Jassem AN. Diagnostic Yield of Laboratory Methods and Value of Viral Genotyping during an Outbreak of Mumps in a Partially Vaccinated Population in British Columbia, Canada. J Clin Microbiol 2018; 56.
  • Tan KE, Anderson M, Krajden M, Petric M, Mak A, Naus M. Mumps virus detection during an outbreak in a highly unvaccinated population in British Columbia. Can J Public Health 2011; 102:47-50.

 


Footnote

[1] Cell culture medium (minimal essential medium or Hanks’ balanced salt solution) or other sterile isotonic solution (e.g. phosphate buffered saline) can be used. The presence of protein, for example 1% bovine albumin, 0.5% gelatin, or 2% serum, stabilizes the virus. Samples without a source of protein in the medium will lose 90%–99% infectivity within 2 hours at 4°C.

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