Rapid multiplex PCR assay to identify respiratory viral pathogens: moving forward diagnosing the common cold.
Upper respiratory tract infections (URIs) can be a serious burden to the healthcare system. The majority of URIs are viral in etiology, but definitive diagnosis can prove difficult due to frequently overlapping clinical presentations of viral and bacterial infections, and the variable sensitivity, and lengthy turn-around time of viral culture. We tested new automated nested multiplex PCR technology, the FilmArray(®) system, in the TAMC department of clinical investigations, to determine the feasibility of replacing the standard viral culture with a rapid turn-around system. We conducted a feasibility study using a single-blinded comparison study, comparing PCR results with archived viral culture results from a convenience sample of cryopreserved archived nasopharyngeal swabs from acutely ill ED patients who presented with complaints of URI symptoms. A total of 61 archived samples were processed. Viral culture had previously identified 31 positive specimens from these samples. The automated nested multiplex PCR detected 38 positive samples. In total, PCR was 94.5% concordant with the previously positive viral culture results. However, PCR was only 63.4% concordant with the negative viral culture results, owing to PCR detection of 11 additional viral pathogens not recovered on viral culture. The average time to process a sample was 75 minutes. We determined that an automated nested multiplex PCR is a feasible alternative to viral culture in an acute clinical setting. We were able to detect at least 94.5% as many viral pathogens as viral culture is able to identify, with a faster turn-around time.
Tripler Army Medical Center, Honolulu, HI.
Clifton P
CP
Layman
Sarah M
SM
Gordon
Diane U
DU
Elegino-Steffens
Willie
W
Agee
Jason
J
Barnhill
Gunther
G
Hsue
D000256:Adenoviridae / Q000302:isolation & purification; D003587:Cytomegalovirus / Q000302:isolation & purification; D005240:Feasibility Studies; D006801:Humans; D060885:Multiplex Polymerase Chain Reaction; D009305:Nasopharynx / Q000821:virology*; D012328:RNA Viruses / Q000302:isolation & purification; D012141:Respiratory Tract Infections / Q000821:virology*; D014776:Virus Cultivation