Respiratory Viral Panel Testing by Multiplex PCR John J. Rushton, PhD, MBA CLINICAL APPLICATION ORDER CODE: Our laboratory is now offering a multiplex PCR Respiratory Viral Panel (RVP) assay that detects the following viral pathogens: • (A, A H1, A H3, A 2009 H1N1, B) RVPCR • RESPIRATORY SYNCYTIAL (RSV) A & B • PARAINFLUENZA (1, 2, 3) • (hMPV) • RHINOVIRUS (HRV) Quick Facts • ADENOVIRUS (B/E, C) Influenza viruses (A/B) are RNA viruses in the family that continuously n RVP detects 14 respiratory undergo genetic changes that can leave the human population vulnerable to seasonal virus types and subtypes changes. Currently circulating influenza A viruses include subtypes H1N1 (including the simultaneously seasonal 2009 H1N1) and H3N2. Respiratory Syncytial Viruses (A/B) are RNA viruses in the family. n Increased sensitivity over with RSV is common in children and adults. with RSV A are thought to and DFA be more severe than infections with RSV B. Parainfluenza Viruses (1/2/3) are RNA viruses in the paramyxoviridae family. The n Flu A Subtyping including: parainfluenza viruses are also commonly identified causative agents of lower respiratory 2009 H1N1, H1, and H3 tract infections (LRTIs) in children, albeit at a lower frequency than RSV. subtypes Human Metapneumovirus (hMPV) is a member of the same virus family as RSV and PIV and has been identified as an important respiratory pathogen in young children with n Turnaround time 1-3 days further studies confirming hMPV infections in persons of all ages. Rhinovirus (HRV) is a RNA virus in the family. The most common infectious agent of both upper and lower respiratory illness, rhinoviruses can cause severe infection in certain populations. Adenoviruses are a diverse group of non-enveloped DNA viruses. The subgenera B,C, and E are frequently associated with upper infections with high rates in closed populations. CLINICAL BACKGROUND

Respiratory viruses are responsible for a wide range of acute respiratory tract infections including the , influenza, and , and represent the most common cause of acute illness in the U.S. Disease severity can be especially high in the young, the immunocompromised and elderly patients. Diagnosis of respiratory infection by clinical symptoms alone is extremely difficult and often inaccurate due to non-specific symptoms.

Virus Common Symptoms Commonly Infected Demographic Influenza A (Flu A) Influenza A H1 (Flu A H1) Influenza A H3 (Flu A H3) Upper respiratory tract infections (URTIs) with All ages, 5-20% of US population Influenza A 2009 H1N1 (2009 H1N1) Influenza B (Flu B) Respiratory Syncytial Virus A (RSV A) LRTIs in infants Infants, children, older adults Respiratory Syncytial Virus B (RSV B) *RSVA thought to be more severe than RSVB Human Metapneumovirus Infants, children Parainfluenza Virus 1 (PIV 1) Laryngotracheobronchitis (croup) Infants, children Parainfluenza Virus 2 (PIV 2) Laryngotracheobronchitis (croup) Infants, children LAB TEST CONNECT Parainfluenza Virus 3 (PIV 3) Bronchiolitis and Infants, children, immunocompromised Rhinovirus (HRV) Mild URTIs and severe LRTIs in at-risk populations All ages Adenovirus (B/E) URTIs All ages, immunocompromised Adenovirus (C) www.tricitieslab.com Respiratory Viral Panel Testing by Multiplex PCR

RESULT INTERPRETATION TEST INFORMATION Detection of the 14 respiratory viral targets listed occurs RESPIRATORY VIRAL PANEL TESTING simultaneously in a single reaction cartridge using a solid-phase BY MULTIPLEX PCR electrochemical detection methodology. Multiplex PCR -RVP Description Respiratory Viral Panel PCR provides a qualitative result based upon the presence (Positive) or absence (Target not Detected) of the viruses contained within the Method PCR panel. Negative results do not preclude respiratory viral infection Order Code RVPCR and should not be used as the sole basis for diagnosis, treatment, or CPT Code 87633 other patient management decisions. Because co-infections occur approximately 10%-30% of the time, there may be more than one Specimen Requirements Flocked nasopharyngeal (NP) virus detected and reported. swab preferred; polyester, rayon or nylon tipped swabs acceptable. Submit in viral METHODOLOGY COMPARISON transport media. Alternate Specimens Bronchoalveolar Lavage (BAL), Diagnostic Testing: Culture / DFA / PCR bronchial washes, and throat DIRECT swabs in viral transport media. MULTIPLEX VIRAL FLUORESCENT PCR CULTURE Comments Qualitative assay reports as ANTIBODY (DFA) Detected or Not Detected SENSITIVITY 91-100% 69-98% 40-80% Schedule Monday - Friday SUBTYPING (Influenza A, RSV, YES NO NO Turnaround time 1-3 days Adeno only) TURNAROUND 1-3 Days 7-14 Days 1 Day TIME # OF VIRUSES 14 7 8 DETECTED DETECT YES NO NO CO-INFECTIONS

Multiplex Polymerase Chain Reaction (PCR) detection offers a highly SELECTED REFERENCES sensitive and specific method of respiratory virus detection and is 1. Gharabaghi et. al Clin Micro and Infection 17(12):1900-06 quickly replacing culture and DFA as the gold standard. In addition to sensitivity and specificity, benefits of multiplex PCR testing 2. Popowitch et. al J Clin Mircobiol 2013 51(5):1528-33 include the ability to obtain the results in hours and demonstrated 3. Virol J. 2013 Jun 7;10:184. doi: 10.1186/1743-422X-10-184.”A cost ability to detect more than one viral infection (co-infection) in a effective real-time PCR for the detection of adenovirus from viral swabs.” Al-Siyabi T, Binkhamis K, Wilcox M, Wong S, Pabbaraju K, Tellier R, Hatchette patient specimen. Emerging evidence suggests patients with viral TF, LeBlanc JJ. co-infections have increased disease severity and more complex clinical management.

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