Materials and Methods for Specimen Collection, Storage, and Shipment
Samples for serologic testing
Serology (serum) samples
The first (acute-phase) serum sample should be collected as soon as possible upon suspicion of mumps disease. Collect 7–10 ml of blood in a red-top or serum-separator tube (SST). Convalescent-phase serum samples should be collected about 2–3 weeks after the acute-phase sample.
Store specimens at 4°C and ship on wet ice packs.
Samples for viral detection
Oral or buccal swab samples
Collect oral or buccal swab samples as soon as mumps disease is suspected. Samples collected when the patient first presents with symptoms have the best chance of having a positive result by RT-PCR.
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). 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 samples have not been as useful as buccal and oral specimens for virus isolation or detection of mumps RNA. Unlike buccal and oral specimens, urine samples may not be positive for mumps virus until >4 days after symptom onset. 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.
 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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