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Aug 19, 2021Print this page

Enterovirus Test Kit: How to Test for Enterovirus D68

Enterovirus D68 Test Kit

Enterovirus D68 (EV-D68), first detected in 1962, is one of more than 100 non-polio enteroviruses. But testing for the Enterovirus still remains challenging for some healthcare facilities.

Unfortunatelytesting for EV-D68 has proven difficult because although on-the-spot tests can detect the presence of an enterovirus, many cannot perform the typing analysis necessary to attribute the virus to a particular strain. Additionally, it’s not always clear when to test for enterovirus and when to attribute symptoms to other respiratory illnesses. 

What are Some Facts About Enterovirus D68?

The Enterovirus Foundation provides the following quick facts about the D68 variation of the Enterovirus:

  • Enterovirus 68, also known as Enterovirus D68 or EV-D68, in one of the more that 100 non-polio enteroviruses and was first identified in California in 1962.
  • Enterovirus 68 has been associated with paralysis and is being investigated as a cause like polio symptoms.
  • Symptoms include: Mild symptoms include runny noses, sneezing, cough, body aches, fever and muscle aches. Severe symptoms include respiratory illness, wheezing and difficulty breathing.
  • Since EV-D68 typically impacts respiratory functions, infected persons spread the virus through coughing, sneezing, or touching surfaces that are then touched by others.

In general, infants, children and teenagers are at highest risk to EV-D68 and become ill most often.  Children with asthma can be at higher risk as well to respiratory illness caused by Enteroviruses.

When to Test for Enterovirus D68

Doctors note that children with asthma or other breathing problems are more likely to develop serious symptoms, and that wheezing coupled with traditional cold and flu symptoms is cause for concern.

Whether you’re a parent, a physician, or both, if a child or patient has expressed any difficulty breathing or is wheezing, along with other symptoms of respiratory illness, it’s a good idea to have them tested for enterovirus D68.

How to Test for Enterovirus D68

So how can healthcare professionals test for suspected enterovirus 68 infection? The CDC and some state health departments can perform enterovirus typing.

If a patient is exhibiting symptoms beyond those of a mild respiratory infection, physicians can take a sample to be sent out for testing, identification, and recording. For enterovirus typing, the CDC will use real-time or nested PCR testing.

Like most respiratory illnesses, nasopharyngeal (NP) swabs and oropharyngeal (OP) swabs are the preferred method of specimen collection for testing, though stool samples and rectal swabbing are also appropriate.

Steps to Collect a Nasopharyngeal (NP) Enterovirus D68 Specimen

  1. Wash hands thoroughly
  2. Put on gloves, surgical mask and protective eye-wear
  3. Choose a swab with a flexible plastic handle and flock tip, such as Puritan’s 25-3317-U.
  4. Insert swab into one nostril straight back (not upwards) and horizontally to the nasopharynx up to the measured distance on the swab handle.
  5. Rotate the swab up to 5 times and hold in place for 5-10 seconds to collect sample material.
  6. Remove swab and insert into a vial containing 1-3ml of viral transport media.
  7. Break the swab handle at scored breakpoint line.
  8. Label the vial with appropriate patient information.

Steps to Collect a Enterovirus D68 Throat Swab Specimen

  1. Wash hands thoroughly
  2. Put on gloves, surgical mask and protective eye-wear
  3. Have the patient facing a strong light to ensure the area to be swabbed is visible
  4. Open the package containing the HydraFlock® flocked swab and aseptically remove swab from package
  5. Have the patient tilt their head backwards, open their mouth, and stick out their tongue
  6. Use a wooden tongue depressor to hold the tongue in place
  7. Without touching the sides of the mouth, use the sterile HydraFlock® flocked swab to swab the the posterior nasopharynx and the tonsillar arches
  8. Insert swab into sterile liquid amies transport system vial
  9. Break the swab handle at scored breakpoint line
  10. Replace cap and tighten to secure
  11. Remove and aseptically dispose of gloves and mask
  12. Label liquid amies vial with appropriate information
  13. Deliver samples to laboratory for testing

How to Submit an Enterovirus Virus Sample for Nested PCR Typing

To submit an NP or OP swab sample to the CDC for enterovirus testing, ship a minimum of 1 ml of specimen in viral transport medium. Freeze at (-20) degrees Celsius and pack for shipping on dry ice.

Our UniTranz-RT™ Transport System is appropriate for the preservation and transport of enterovirus 68. You can choose our viral transport medium system on its own, with flocked swabs (for superior collection and elution of specimen), or with polyester tipped swabs. The systems are packaged as a unit to make sampling, preserving, and transporting specimen as simple as possible.

All submitted specimens must include CDC form 50.34 and a patient summary form for each patient for whome specimens are being submitted. Finally, before sending specimens, the CDC asks that proper infection control measures are followed.

How to Help Your Patients Now

As a healthcare professional, you’ll likely encounter a number of anxious parents. Some children diagnosed with this year’s EV-D68 have experienced unexplained paralysis or extreme muscle weakness in their limbs. A few individuals diagnosed with the virus have died; though it’s unclear whether all the deaths can be directly attributed to EV-D68.

But before parents go encasing their children in plastic bubbles, you can help them maintain a realistic perspective by reminding them of the following information:

1) Not Every Case of EV-D68 is Serious

Many people will contract the virus and experience little more than mild flu-like symptoms including: fever, stuffy nose, cough, and muscle aches. “Few people who contract EV-D68 develop symptoms other than a runny nose and a cough," said Dr. Andi Shane, associate professor of pediatrics and global health at Emory University's School of Medicine and Children's Healthcare of Atlanta. 

2) The Impact of EV-D68 Will Be Much Smaller than the Flu

The number of cases of enterovirus D68 will pale in comparison to the number of cases of the flu, which can kill hundreds of children in a given season. (Reassuring, no?). This might be a good opportunity to remind parents about the importance of getting their children (and themselves) vaccinated for the flu. 

3) Hand Washing is the Best Protection

There is currently no specific treatment for EV-D68, so prevention is still the best medicine. In addition to collecting samples and reminding them of the relative rarity of the virus, you can help concerned parents by demonstrating (and having their children participate in) proper hand-washing procedures and by stressing the importance of good hand hygiene for staving off illness.

Want more information about using Puritan swabs or transport systems to test for viruses this cold and flu season?



Fox, Maggie. Why is Enterovirus D68 Such a Problem for Kids This Year? CDC Looking. NBC News. Retrieved October 9, 2014 from: http://www.nbcnews.com/health/health-news/why-enterovirus-d68-such-problem-kids-year-cdc-looking-n221461

Goodman, Brenda. Reviewed by Bhargava, Hansa, MD. Enterovirus D68: What Parents Need to Know. Cold, Flu, & Cough Health Center. September 9, 2014. Updated October 6, 2014. Retrieved October 9, 2014 from: http://www.webmd.com/cold-and-flu/news/20140909/enterovirus-d68-parents

Specimen Collection, Storage, & Shipment. Non-Polio Enterovirus. Centers for Disease Control and Prevention. Retrieved October 9, 2014 from: http://www.cdc.gov/non-polio-enterovirus/lab-testing/specimen-collection.html

St. James, Janet. Enterovirus testing frustrating, doctor says. September 10, 2014. Retrieved October 9, 2014 from: http://www.usatoday.com/story/news/nation/2014/09/10/enterovirus-testing-frustrating-doctor-says/15373675/

Enterovirus D68 for Health Care Professionals. Non-Polio Enterovirus. Centers for Disease Control and Prevention. October 1, 2014. Retrieved October 9, 2014 from: http://www.cdc.gov/non-polio-enterovirus/hcp/EV-D68-hcp.html

Topics: Flocked Swabs, Media Transport Systems, Diagnostics, Specimen Collection Procedure


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