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Enterovirus 68 Testing

Submitting Samples for Enterovirus 68 Testing through CDC

The Centers for Disease Control and Prevention (CDC) currently requires prior approval for testing and typing of Enterovirus 68 (EV-D68) specimens. Presently the CDC is prioritizing samples from patients with severe respiratory illness who are known to be positive for rhinovirus (RV)/enterovirus (EV). For EV-D68 testing, the CDC prefers a prior RV/EV positive test. Specimens should be limited to those from patients with severe illness (for example, patients in the PICU).

Please contact your Local Health Department or the Oregon Acute and Communicable Disease Program (971-673-1111) if you would like to submit specimens for EV-D68 testing to the CDC.


Specimen Collection Information

Timing of Specimen Collection

Collecting specimens during the first week of illness is ideal although the virus can be shed in stool for several weeks. A specimen set collected in the second week of illness should include a rectal swab or stool sample.

NP and OP swabs have had a high diagnostic yield. Stool or rectal swabs may also be submitted. Please submit each specimen in a separate container.

Specimen collection and handling, by specimen type

Specimen Type Specimen Collection Storage Transport Test Type Comments

CSF

Collect in sterile container, no special medium required.

Volume: >150 ul

Freeze at -20 degrees C

Ship on dry ice

Real-time PCR (generic)

Nested PCR (typing)

Collect within 0-4 days of symptom onset (the earlier the better)

Serum*

Collect in sterile container with no anticoagulants

Volume: >150 ul

Freeze at -20 degrees C

Ship on dry ice

Real-time PCR (generic)

Nested PCR (typing)

No serology is done. Collect early in infection from younger patients.

Respiratory**

For NP/OP washes and aspirates, collect in sterile container, no special medium required.

For NP/OP swabs, store in viral transport medium.

Minimum volume: 2ml

Freeze at -20 degrees C

Ship on dry ice

Real-time PCR (generic)

Nested PCR (typing)

NP and OP swabs have a high diagnostic yield.
Stool

Collect in sterile container, no special medium required.

For rectal swab, store in viral transport medium.

Volume: >1 gram

Freeze at -20 degrees C

Ship on dry ice

Real-time PCR (generic)

Nested PCR (typing)

Virus present for longer than other specimen types

Submit each specimen in a separate container.

Fresh-frozen tissue (preferred) Placed directly on dry ice or liquid nitrogen.

Freeze at -70 degrees C

Ship on dry ice

Real-time PCR (generic)

Nested PCR (typing)

Formalin-fixed or formalin-fixed, paraffin-embedded tissue

Avoid prolonged fixation - can fix in formalin then transfer to 100% ethanol. n/a Ship at room temperature with paraffin blocks in carries to prevent breakage.

Real-time PCR (generic)

Nested PCR (typing)

Acceptable specimen type, but lower sensitivity.

*Notes for Serum Specimens

  • For each serum specimen, collect (adults and children >6 kg: 5 ml, children <6k g: 2 ml) of whole blood into a serum separator tube (marble- or tiger-top SST). A minimum of 1 ml of whole blood is needed for testing of pediatric patients. Allow to clot at room temperature for a minimum of 30 minute and centrifuge.

**Comments Regarding Acceptable Swabs for Respiratory Specimens

  • For nasopharyngeal (NP), oropharyngeal (OP), and/or rectal swabs, please use only sterile dacron or rayon swabs with plastic shafts. Alternatively, you may use flocked swabs if available.
  • DO NOT use calcium alginate swabs or swabs with wooden sticks, as they may contain substances that inactivate some viruses and inhibit some molecular assays.
  • Place the NP/OP swabs immediately into a sterile vial containing 2 ml of viral transport media without antibiotics.
  • Aseptically, cut or break applicator stick off near the tip to permit tightening of the cap.
  • Sterile PBS or Hank’s balanced salt solution (HBSS) (no antibiotics) can be used in lieu of viral transport medium.
  • The table above reflects the most recent guidance provided to the Public Health Division by the CDC and may differ from guidance posted on the CDC's webpages. 

Shipment

Please ship samples through the Oregon State Public Health Laboratory.


Additional Information

For additional resources regarding enterovirus 68, including disease reporting requirements, guidelines for providers, and additional links to the CDC, visit the Public Health Division's Emerging Respiratory Infections webpage.

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