Appendix 15-1 

Collection of Specimens for

Typing of Rubella Virus 

Please coordinate the shipping of specimens with Dr. Joseph Icenogle’s office (see 

address below). 

I. Respiratory Specimens 

Throat swab or nasopharyngeal washes 

Materials: 

● sterile swabs 

● 3 ml aliquots of viral transport medium (VTM: sterile PBS or suitable isotonic solution 

such as Hanks BSS, containing antibiotics (100 units/ml penicillin, 100 μg/ml streptomycin) 

and either 2 % fetal bovine serum or 0.5% gelatin in 15 ml polycarbonate or polystyrene 

centrifuge tubes 

● 5 ml plastic syringes 

● plastic aspirators or 30 ml syringe 

● Styrofoam shipping containers 

Instructions: 

1. Attempt to obtain the sample as soon as possible after onset of rash but a maximum of 4 

days after rash onset. 

2. A throat swab as if you were obtaining a bacterial culture is the preferred method. 

Alternative: An alternative method for specimen collection is a nasal wash 

(nasopharyngeal aspirate) using a syringe attached to a small, plastic tube and 3–5 ml of 

VTM. After placing VTM in the nose, aspirate as much of the material as possible and rinse 

the tube with a small volume (2ml) of VTM. 

OR 

Alternatively, sterile swabs can be used to wipe the nose and throat. Place both swabs in 

a tube containing 2–3 ml of transport medium. The virus is extremely cell-associated, so 

attempt to swab the throat and nasal passages to collect epithelial cells. 

3. Procedures for shipping of specimens 

Preferred method: Keep all specimens on wet ice or at 4° C and ship as soon as possible 

on wet ice (see address below). 

Other acceptable methods: If immediate, cold shipment (within 48 hrs) cannot be 

arranged or is not convenient. Nose and throat swabs can be removed from the transport 

medium after allowing some time for elution of virus (at least 1 hr). Nasal wash specimens 

can be centrifuged at 2500 x g for 15 minutes at 4° C and the pellet resuspended in 1 ml 

of tissue culture medium. If possible, the supernatant can be saved in a separate tube. The 

samples should be frozen and shipped at -70° C (dry ice). If centrifugation is not available 

the whole specimen can be frozen (preferably at -70° C) and shipped on dry ice. 

II. Urine specimens 

Methods 

1. First morning voided specimens are ideal, but any urine collection is adequate. Collect up 

to 50–100 ml of urine in a 50 ml centrifuge tube or a urine specimen container. 

2. Centrifuge at 2500 x g for 15 minutes at 4° C to pellet the sediment. Resuspend the 

sediment in 2–3 ml of VTM (see above) or any cell culture medium (DMEM, EMEM, 

RPMI plus antibiotics) and ship. Preferably specimens that have been centrifuged and 

resuspended should be frozen at -70° C and shipped on dry ice. If dry ice is not available, 

however, they can be stored at 4° C and shipped on wet ice. 

CS106190
Appendix 15-2 

III. Shipping 

It is best to send specimens by express courier early in the week (Monday or Tuesday) so the 

shipment can be tracked, if necessary. Before shipping, call or e-mail Dr. Icenogle’s laboratory. 

Ship to: 

Dr. Joseph Icenogle 

Rubella Virus Section, C-22 

DASH Group #81 

1600 Clifton Rd., NE 

Atlanta, GA 30033 USA 

Phone: 404-639-4557 

E-mail: jci1@cdc.gov