Comparison of the Idaho Technology FilmArray System to Real-Time PCR for Detection of Respiratory Pathogens in Children

Abstract

The FilmArray Respiratory Panel (RP) multiplexed nucleic acid amplification test (Idaho Technology, Inc., Salt Lake City, UT) was compared to laboratory-developed real-time PCR assays for the detection of various respiratory viruses and certain bacterial pathogens. A total of 215 frozen archived pediatric respiratory specimens previously characterized as either negative or positive for one or more pathogens by real-time PCR were examined using the FilmArray RP system. Overall agreement between the FilmArray RP and corresponding real-time PCR assays for shared analytes was 98.6% (kappa ‫؍‬ 0.92 [95% confidence interval (CI), 0.89 to 0.94]). The combined positive percent agreement was 89.4% (95% CI, 85.4 to 92.6); the negative percent agreement was 99.6% (95% CI, 99.2 to 99.8). The mean real-time PCR threshold cycle (C T ) value for specimens with discordant results was 36.46 ؎ 4.54. Detection of coinfections and correct identification of influenza A virus subtypes were comparable to those of realtime PCR when using the FilmArray RP. The greatest comparative difference in sensitivity was observed for adenovirus; only 11 of 24 (45.8%; 95% CI, 27.9 to 64.9) clinical specimens positive for adenovirus by real-time PCR were also positive by the FilmArray RP. In addition, upon testing 20 characterized adenovirus serotypes prepared at high and low viral loads, the FilmArray RP did not detect serotypes 6 and 41 at either level and failed to detect serotypes 2, 20, 35, and 37 when viral loads were low. The FilmArray RP system is rapid and extremely user-friendly, with results available in just over 1 h with almost no labor involved. Its low throughput is a significant drawback for laboratories receiving large numbers of specimens, as only a single sample can be processed at a time with one instrument.A variety of viruses and bacteria are responsible for acute upper and lower respiratory tract infections in children and adults worldwide. Severe and even fatal disease can occur in the very young, the elderly, the immunocompromised, and those individuals with underlying conditions that affect cardiopulmonary function. While certain clinical syndromes are classically associated with specific pathogens, in reality, the syndromes caused by these organisms are often indistinguishable, and rapid and accurate detection is important for appropriate patient management and prevention of nosocomial spread (2,5,6,7,13,23,26,31,35). Through the years, PCR has shown excellent clinical utility over the more traditional laboratory methods of virus culture and direct antigen tests for the sensitive detection of respiratory viruses (8,15,24,25,34), and both laboratory-developed and commercial molecular assays are now available (22). However, because of the complexity of these molecular tests, implementation has been a challenge for the routine clinical laboratory.More recently, significant advances in microfluidics, microelectronics, and microfabrication have paved the way for the development of simplified molecular systems with the possibili...

Comparison of three multiplex PCR assays for detection of respiratory viruses: Anyplex II RV16, AdvanSure RV, and Real‐Q RV

“…These findings suggest that the discordant results were frequently reported in samples containing low viral copy. Other studies also indicated that discrepant results between RV assay was quite associated with sample's viral load 7, 8, 19, 20, 21. One samples by AP and three samples by AD identified as INF A positive by additional testing.…”

“…Performance evaluation studies for newly developed multiplex RV kits are weak, do not establish the reference standard method and therefore do not sufficiently calculate sensitivity and specificity of each test. Instead, some studies have suggested the reference test with in‐house multiplex real‐time PCR or commercial duplex PCR tests 5, 6, 7, 8. We performed a direct comparison of three commercial multiplex assays and produced the values of the agreement and kappa instead of sensitivity and specificity with the reference tests.…”

Severe Complications in Influenza-like Illnesses

“…However, it is less likely that a listed tested virus was present and missed, as the PCR testing has .92% sensitivity for each tested virus, except certain adenovirus serotypes. 17 It should be noted that severe complications were not significantly more common among children without virus identified. Additionally, although we were able to perform subanalyses for pandemic H1N1 infection, the analyses were limited because of sample size.…”

“…17 Specific viruses identified included influenza A and B, parainfluenza viruses, rhinovirus, human metapneumovirus, adenovirus, and respiratory syncytial virus (RSV). Results of multiplex viral respiratory PCR testing were collected after the ED visit was complete.…”

Disseminated Adenovirus Infection in Cancer Patients Presenting with Focal Pulmonary Consolidation

“…In the study by Pierce et al, the sensitivity of Filmarray RP version 1.6 was 46% compared to standard individual real-time quantitative PCR assays (13,14). Among the currently FDA-approved molecular assays that detect more than 3 respiratory pathogens, the sensitivity for detection of ADV ranges from 57.1% to 100%; with a sensitivity of 57.1% reported for FilmArray RP v1.6 (13,14). The low sensitivity of FilmArray RP v1.6 for ADV specifically relates to ADV C, serotypes 2 and 6 (13).…”