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Non-variola Orthopoxvirus

PCR Testing for Non-variola Orthopoxvirus.

1. Dry swab of lesion is preferred.

2. Vesicle fluid, skin, crust, “roof”

1. Swab – Swab material should be synthetic, i.e., rayon, polyester, or Dacron. Calcium alginate or charcoal impregnated swabs should not be used, nor should wood-shaft swabs.
2. 1.5- or 2-mL sterile, screw-capped tube with O-ring or 15-mL sterile, screw- capped tube.
3. For collection of vesicle fluid, a 26 or 27-gauge needle attached to a tuberculin syringe is necessary.
4. Cold packs or dry ice.
5. Shipping boxes/containers with appropriate shipping labels (Category B Infectious Substance).

1. Don personal protective equipment per CDC guidance.

2. Be sure to properly label the specimen tube with the patient’s name, date of birth, date of collection, and location of the swabbed lesion. If there is not enough space on the tube to clearly write the location of the swabbed lesion (i.e., left forearm), write the location clearly on the outside of the biohazard specimen bag.

3. Special Instructions for Specimen Collection:

  • Vigorously swab or brush lesion with two separate sterile dry polyester or Dacron swabs.
  • Insert the swab, swab-end down, into separate 1.5- or 2-mL screw- capped tube with an O-ring or place the entire swab in a separate sterile container (i.e., 15-mL sterile, screw-capped tube).
  • Break off the swabs at the scored line on each of the swab shafts. If the swab does not have a scored line, please cut the swab shaft to fit in the screw-capped tube.
  • Tightly cap the tubes for transport.
  • If the patient presents with more than one lesion, please repeat the collection steps for a second lesion, preferably on a different part of the body and/or with a different physical presentation.
  • If two sets of specimens are collected, please place these in SEPARATE biohazard specimen bags. Mark the ‘Refrigerated’ box on the specimen bag.

4. After collection, all specimens should be stored at refrigerated temperature (2- 8°C) until shipped. If longer storage is required, specimens can be frozen.

5. Specimens should be submitted through the Biothreat page on LimsNet, an online system that will make results available as PDF files after they are released at the lab. Each set of lesion swabs (2 swabs per one lesion) should be submitted separately in LimsNet, creating up to two LimsNet submissions per one patient.

  • To get a free LimsNet account established at your facility for electronic submission and results reporting, call the help desk at (317) 921-5506 or email to

6. If you have questions, please contact the IDOHL BT-Clinical Microbiology and Environmental Virology Division Director at 317-921-5842 or the Virology & Serology Division Director at 317-921-5555.

1. Wrap the labeled specimen container with absorbent material and place in a biohazard specimen bag. Be sure to package each patient’s specimens individually to avoid cross-contamination. Please ship non-variola orthopoxvirus specimens separately from any other specimens coming from the submitting laboratory.

2. Place the requisition form in the side pocket of the biohazard bag. Never place the requisition form in with the specimen in case the specimen leaks during transit. If the specimen bag does not have 2 compartments, place the paperwork in a separate Ziploc bag.

3. Place the specimen(s) in a Styrofoam container with sufficient cold packs to maintain refrigerated temperature during shipment. If needed, dry ice can also be used if the specimen is frozen and/or transport time may be longer than 24 hours. If dry ice is used, do not form an airtight seal on the Styrofoam container because dry ice releases carbon dioxide gas.

4. Place the Styrofoam container into a cardboard shipping box, close lid and seal.

5. Ship or transport by courier, compliant with DOT and IATA regulations (Category B Infectious Substance).

Ship to:
Indiana Department of Health Laboratories: Biothreat Laboratory
550 W. 16th St., Suite B
Indianapolis, IN 46202

1-2 business days.
Specimens may be forwarded to the CDC for characterization testing