Influenza Virus RNA Detection  

IMPORTANT! Pre-approval needed from state or local health department. Local or State Health Department. Send approval to or call (402) 559-9444.

NPHL has moved to an electronic ordering system, NUlirt, and it is now the preferred method to order all tests. Once ordered, print a "batch list" (see NUlirt Guide).

Forms Required - Nulirt batch list or NPHL Test Request Form (completed in entirety) must accompany the specimen to include the following information: symptoms and date of onset, pregnancy status, travel and vaccine history. Testing will not be performed without all required information.

Please call NPHL Client Services at (402) 559-2440 or toll free at (800) 334-0459 for further clarification.


Test: Influenza Virus RNA Detection
Order in NUlirt or contact Robin Williams, DHHS for Supplemental Form  
Synonym: Flu PCR or RT-PCR, Influenza Confirmation, FLUPCR  
Method: Qualitative nucleic acid amplified real time RT-PCR assay  
Availability: Varies according to testing volume, refer to laboratory for testing interval  
Specimen: Upper Respiratory Tract: Posterior-pharyngeal swab, nasopharyngeal or nasal swab, nasal aspirate, or nasal wash.
Lower Respiratory Tract: Tracheal aspirate, bronchoalveolar lavage (BAL), bronchial washes, endotracheal aspirate, endotracheal wash, sputum, lung tissue. 
(contact the laboratory for guidance if other specimen sources are being considered)
Collection Device: Transport human respiratory specimens in Viral Transport Media (VTM)
Specimen Labeling: Test subject to CLIA regulations and required 2 patient identifiers on specimen container and requisition forms  
Volume: Collect 2 - 3 mL from nasopharyngeal wash (using normal saline) or 2 - 3 mL BAL  

Prior to and during shipment:  Refrigerated at 2-8° C. Ship within 72h or freeze.
Shipping instructions, including specimen-handling requirements during transport:   Seal screw-top tubes with Parafilm™ or tape and place in biohazard bag with absorbent material.
Place specimen inside leak proof secondary container (biohazard bag and if transporting by air, use Tyvek™ Envelope) with adsorbent material. The secondary container must be packed inside an approved Category B shipping container filled with frozen gel-packs to keep specimen refrigerated or dry ice to keep specimen frozen. Position NUlirt packing list or NPHL Test Order form between secondary container and box lid. Seal with tape.  Label outer box as UN3373 Category B, Biological Substance. Transport by NPHL ground courier. If courier not available, ship by FedEx, UPS, or USPS (1st Class, Priority or Express Mail). Follow appropriate DOT/IATA Regulation Guidelines.  For assistance, call NPHL State Training Coordinator at (402) 559-3590.  

NUlirt packing list or Influenza Virus RNA Supplemental Form (completed in entirety) must accompany specimens.

Unacceptable: Slides, smears; unacceptable sources; Sputum, tracheal aspirates, or throat swabs or samples collected in non-viral transport media or using a bacterial culture swab; dry swabs; wood shaft or calcium alginate swabs; samples collected in EIA or extracted transport media.  
Specimen Stability: 2-8°C, 72h; Frozen, 1month  
Reference Interval: Negative for Influenza Type A virus, Negative for Influenza virus type B, Influenza A subtype H3 virus (seasonal), Influenza 2009 A (H1N1), Influenza B Yamagata lineage, Influenza B Victoria lineage.  
Reportable Disease: Positive results (influenza A virus [untypable], influenza B virus [untypable], influenza A virus subtype H3 [seasonal], influenza A virus subtype 2009 [H1N1], influenza B Yamagata lineage or influenza B Victoria lineage) are reported within 7d to local or state health department. 
Both positive and negative results are reported to the state within 7d for ELR laboratories only.   

A positive point of care POC kit (antigen or PCR) test that occurs during the “off season” or early-on during the respiratory season should be submitted to NPHL for confirmation testing.

Refer to Nebraska DHHS Title 173, Communicable Diseases. Ship all non-typable influenza A virus specimens to NPHL  
Comments: Pre-approval needed from state or local health department.  Send approval to or call (402) 559-9444.

A false negative result may occur if a specimen is improperly collected, transported, or handled.  False negative results may also occur if amplification inhibitors are present in the specimen or if inadequate numbers of organisms are present in the specimen.

NUlirt packing list or appropriate paperwork (completed in entirety) must accompany specimens.
Revised: 6/11/2021 
If you have questions about proper specimen collection, please call NPHL Client Services at (402) 559-2440 or toll free at (800) 334-0459.