Assay Sheet
Test ID
8100 Seasonal Influenza A H1 and H3 Subtyping RT-PCR
CPT Code
87798 (x2)
Clinical Utility
Influenza A viruses are medically important pathogens that cause significant morbidity and mortality throughout the world. The recent emergence of the 2009 H1N1 influenza A virus has lead to increased vigilance in monitoring these infections. Availability of several antiviral drugs coupled with differing resistance patterns between subtypes has led to a need for diagnostic testing that is able to rapidly and accurately subtype influenza A specimens. Utilizing molecular testing for subtyping enables the clinician to choose appropriate antiviral drug therapy as well as assisting in epidemiological tracking of influenza outbreaks.
Procedure
The assay consists of two real time RT- PCR reactions that allows for differentiation between seasonal influenza A H1 and influenza A H3.
Extraction of influenza A nucleic acid from respiratory specimen is followed by reverse transcription of viral RNA, then amplification and detection of cDNA using real-time PCR. An internal control is added to ensure the extraction was performed correctly and the PCR reaction was not inhibited.
Note: The seasonal influenza A H1 assay will be negative if the patient is infected with 2009 H1N1. If confirmation of 2009 H1N1 infection is needed, ViraCor test code 7700 should be ordered.
Specimen type & specimen handling
Bronchial Lavage/Bronchial Wash: 2 mls collected in a sterile, screw top tube. Ship at ambient temperature Monday thru Friday. Specimen must be received within 96 hrs of collection.
Sputum: 2 mls collected in a sterile container, then transferred to sterile, screw top tube for shipment. Ship at ambient temperature Monday thru Friday. Specimen must be received within 96 hrs of collection.
Throat Gargle: 2 mls collected in a sterile container then transferred to sterile, screw top tube for shipment. Ship at ambient temperature Monday thru Friday. Specimen must be received within 96 hrs of collection.
Upper respiratory aspirate (NP aspirate, nasal aspirate/wash, tracheal aspirate, etc.): 2 mls collected in a sterile, screw top tube. Ship at ambient temperature Monday thru Friday. Specimen must be received within 96 hrs of collection.
Upper respiratory swab (NP swab, throat swab): Sterile swab placed in 2 ml sterile saline, M4, or viral transport media in a sterile, screw top tube. Do not use calcium alginate swab or wood shafted swab. Ship at ambient temperature Monday thru Friday. Specimen must be received within 96 hrs of collection.
All suction-type collection devices are inappropriate for specimen transport. Transfer specimen into sterile, leakproof tube for transport.
Call ViraCor for authorization prior to sending any specimen type other than those listed above.
If another specimen type has received authorization for testing the following comment will appear in the final report: "The clinical utility of this result has not yet been demonstrated in the peer reviewed literature and is therefore unknown."
Causes for Rejection
Wood shafted swab, calcium alginate swab
Call ViraCor at 800-305-5198 if specimen is greater than 96 hrs old
Specimen types other than those listed above that were sent without prior authorization
Specificity
The primers and probes used in this assay are specific for influenza A seasonal H1 and influenza A H3, respectively, based on similarity search algorithms. For the influenza A seasonal H1 portion of the assay, no cross reactivity was detected when tested against influenza A 2009 H1N1, influenza A H3, influenza B, RSV A, RSV B, parainfluenza 1, parainfluenza 2, parainfluenza 3, hMPV, rhinovirus, and adenovirus. Additionally, no cross reactivity was detected for the influenza A H3 portion of the assay when tested against influenza A seasonal H1, influenza A 2009 H1N1, influenza B, RSV A, RSV B, parainfluenza 1, parainfluenza 2, parainfluenza 3, hMPV, rhinovirus, and adenovirus.
Influenza A Seasonal h1 and influenza a h3 real time rt-pcr assay range
Limit of Detection (LOD): 500 copies/ml
Qualitative results (Positive/Not Detected) for both influenza A seasonal H1 and influenza A H3
Turnaround Time
Same day (within 8 to 12 hours of receiving specimen), Monday through Saturday
Reflex Testing Options Available
ViraCor 2009 H1N1 Influenza A Real-Time RT-PCR (7700) Reflex to Seasonal Influenza A H1 & H3 Subtyping (8100)
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8400
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Same-day results for both tests.
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If 7700 result is Influenza A positive and 2009 H1N1 negative, then reflex to 8100
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Note: Go to www.viracor.com for additional respiratory virus reflex testing options.
Shipping
Ship Monday through Friday. Friday shipments must be labeled for Saturday delivery. All specimens must be labeled with patient's name and collection date. Multiple tests can be run on one specimen. Ship specimens FedEx Priority Overnight® to:
ViraCor Laboratories, 1001 NW Technology Dr, Lee's Summit, MO 64086
The CPT codes provided are based on ViraCor's interpretation of the American Medical Association's Current Procedural Terminology (CPT) codes and are provided for informational purposes only. CPT coding is the sole responsibility of the billing party. Questions regarding coding should be addressed to your local Medicare carrier. ViraCor assumes no responsibility for billing errors due to reliance on the CPT codes illustrated in this material. PCR tests are performed pursuant to a license agreement with Roche Molecular Systems, Inc. This assay was developed and the performance characteristics were determined at ViraCor Laboratories. This test is performed in a CLIA certified laboratory. FDA approval is not required for the performance of this test.
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