Smallpox Specimen Collection

Materials needed

• 2—Sterile screw-capped plastic tube with O-ring (1.5 to 2 mL)
• 2—Sterile dry polyester or Dacron swabs

Procedure

1. Swab or brush posterior tonsillar tissue with a sterile dry polyester or Dacron swab.
2. Break off end of applicator into a 1.5 or 2 mL screw-capped tube with O-ring or place entire swab in a sterile container. DO NOT ADD ANY VIRAL TRANSPORT MEDIA.

Materials needed

• 2—Sterile screw-capped plastic tube with O-ring (1.5 to 2 mL)
• 2—Sterile dry polyester or Dacron swabs

Procedure

1. Swab the nasopharynx with a sterile dry polyester or Dacron swab.
2. Break off end of applicator into a 1.5 or 2 mL screw-capped tube with O-ring or place entire swab in a sterile container. DO NOT ADD ANY VIRAL TRANSPORT MEDIA.

Materials needed

• 1—Punch biopsy kit (2.5 mm for pediatrics; 3.5 or 4 mm for adults)
• 1—Needle driver
• 1—Sutures
• 1—Suture removal kit
• 1—Container of 10% neutral buffered formalin
• 2—Sterile screw-capped plastic tube with O-ring (1.5 to 2 mL)
• Multiple—Alcohol wipes

Procedure

1. Use appropriate sterile technique and skin sanitation.
2. Biopsy 2 lesions with 3.5 or 4 mm biopsy punch (2.5 mm for pediatrics).
3. Place one biopsy specimen in formalin.
4. Place one biopsy specimen in a 1.5 to 2 mL screw-capped plastic vial with O-ring. DO NOT ADD ANY VIRAL TRANSPORT MEDIA.

Lesion fluid can be collected by swab, smear or touch prep slides. In addition, if electron microscopy materials are available this type of specimen collection can also be used.

Materials needed

• 2—Disposable scalpel with no. 10 blade, or
• 2—26 Gauge needle
• 4—Sterile screw-capped plastic vials with O-ring (1.5 to 2 mL)
• 4–8—Sterile dry polyester or Dacron swabs
• Multiple—Alcohol wipes

Procedure

1. Sanitize lesion with an alcohol wipe, allow to dry.
2. Use a disposable scalpel (or a sterile 26 Gauge needle) to open and remove the top of the vesicle or pustule (do not send the scalpel or needle). Retain lesion roof for testing.
3. Swab the base of the lesion with a sterile polyester or Dacron swab.
4. Break off end of applicator into a 1.5 or 2 mL screw-capped tube with O-ring or place entire swab in a sterile container. DO NOT ADD ANY VIRAL TRANSPORT MEDIA.

Materials needed

• 2—Disposable scalpel with no. 10 blade, or
• 2—26 Gauge needle
• 4—Clean plastic or glass microscope slides
• 4—Plastic single-slide holders
• 4–8—Sterile dry polyester or Dacron swabs (if needed)
• Multiple—Alcohol wipes
• Parafilm (optional)

Procedure

1. Sanitize lesion with an alcohol wipe, allow to dry.
2. Use a disposable scalpel (or a sterile 26 Gauge needle) to open and remove the top of the vesicle or pustule (do not send the scalpel or needle). Retain lesion roof for testing.
3. Scrape the base of the vesicle or pustule with the blunt edge of the scalpel, or with the end of an applicator stick or swab.
4. Smear the scrapings onto a clean microscope slide.
5. Apply a microscope slide to the vesicular or pustular fluid multiple times, with progressive movement of the slide, to make a touch prep.
6. Allow slides and grids to air dry for approximately 10 minutes.
7. Store slides from different patients in separate plastic slide holders to prevent cross-contamination (parafilm may be used to wrap the slide holder to prevent accidental opening).

Materials needed

• 2—Disposable scalpel with no. 10 blade, or
• 2—26 Gauge needle
• 2–4—Formvar/carbon-coated mesh electron microscopy grids
• 1—Electron microscopy quality forceps
• 1—Electron microscopy grid box
• Multiple—Alcohol wipes

Procedure

1. Sanitize lesion with an alcohol wipe, allow to dry.
2. Use a disposable scalpel (or a sterile 26 Gauge needle) to open and remove the top of the vesicle or pustule (do not send the scalpel or needle). Retain lesion roof for testing.
3. Lightly touch the "shiny side" of an electron microscope grid to the unroofed base of the lesion.
4. Repeat this procedure two more times, varying the pressure applied to the unroofed lesion (lighter or firmer pressure).
5. Place in grid box and record which slot is used for each patient specimen.

Materials needed

• 2—Disposable scalpel with no. 10 blade, or
• 2—26 Gauge needle
• 4—Sterile screw-capped plastic vials with O-ring (1.5 to 2 mL)
• Multiple— Alcohol wipes

Procedure

1. Sanitize lesion with an alcohol wipe, allow to dry.
2. Use a disposable scalpel (or a sterile 26 Gauge needle) to open and remove the top of the vesicle or pustule (do not send the scalpel or needle).
3. Place the skin of the vesicle top into a 1.5 to 2 mL sterile screw-capped plastic tube with O-ring. DO NOT ADD ANY VIRAL TRANSPORT MEDIA.

Materials needed

• 1—26 Gauge needle
• 2—Sterile screw-capped plastic vials with O-ring (1.5 to 2 mL)
• Multiple—Alcohol wipes

Procedure

1. Sanitize skin with an alcohol wipe, allow to dry.
2. Use a 26 Gauge needle to pick or dislodge at least 4 scabs; two scabs each from at least two body locations.
3. Place scabs from each location in separate sterile O-ring vials.

Materials needed

• 1—5 or 10 cc syringe with needle
• 1—Vacutainer holder
• 2—Vacutainer needles
• 1—10 cc red/gray, gold, or red-topped serum separator tube for serum collection
• 1—Lavender-topped tube (potassium EDTA) for whole blood collection

Blood collection procedure

1. Collect 7 to 10 cc of patient blood into a red/gray (marbled), gold, or red topped serum separator tube when patient is first identified.
2. Spin tubes to separate serum.
3. Save the serum in at least 2 aliquots, 1 for immediate testing and the other for paired sera testing at the convalescent–stage of disease.
4. Collect 3 to 5 cc of whole blood into a lavender-topped tube.
5. Gently invert the tube to mix the blood with the anticoagulant.
6. Obtain convalescent-phase serum 4 to 6 weeks after initial acute-phase serum collection.
7. Send convalescent-phase serum with the remaining acute-phase serum aliquot.