Comparison of the GenMark Diagnostics eSensor Respiratory Viral Panel to Real-Time PCR for Detection of Respiratory Viruses in Children

Abstract

A novel eSensor respiratory viral panel (eSensor RVP) multiplexed nucleic acid amplification test (GenMark Diagnostics, Inc., Carlsbad, CA) was compared to laboratory-developed real-time PCR assays for the detection of various respiratory viruses. A total of 250 frozen archived pediatric respiratory specimens previously characterized as either negative or positive for one or more viruses by real-time PCR were examined using the eSensor RVP. Overall agreement between the eSensor RVP and corresponding real-time PCR assays for shared analytes was 99.2% (kappa ‫؍‬ 0.96 [95% confidence interval {CI}, 0.94 to 0.98]). The combined positive percent agreement was 95.4% (95% CI, 92.5 to 97.3); the negative percent agreement was 99.7% (95% CI, 99.4 to 99.8). The mean real-time PCR threshold cycle (C T ) value for specimens with discordant results was 39.73 (95% CI, 38.03 to 41.43). Detection of coinfections and correct identification of influenza A virus subtypes were comparable between methods. Of note, the eSensor RVP rhinovirus assay was found to be more sensitive and specific than the corresponding rhinovirus real-time PCR. In contrast, the eSensor RVP adenovirus B, C, and E assays demonstrated some cross-reactivity when tested against known adenovirus serotypes representing groups A through F. The eSensor RVP is robust and relatively easy to perform, it involves a unique biosensor technology for target detection, and its multiplexed design allows for efficient and simultaneous interrogation of a single specimen for multiple viruses. Potential drawbacks include a slower turnaround time and the need to manipulate amplified product during the protocol, increasing the possibility of contamination.A cute viral upper and lower respiratory tract infections are responsible for substantial morbidity in both pediatric and adult populations worldwide. These infections can be severe and even fatal, especially in susceptible infants, older adults, patients with compromised immune systems, and individuals with underlying cardiopulmonary diseases. Rapid and accurate laboratory detection of respiratory viruses plays an important role in clinical management, guiding the appropriate prescription of antivirals, reducing the need for additional diagnostic studies and hospital procedures, and limiting the use of unnecessary antibiotics (2,4,5,6,10,19,31,35). In addition, a virologic diagnosis is instrumental to efforts focused on prevention of health care-associated infections (3,9,23,24).PCR has demonstrated superior sensitivity for the detection of respiratory viruses over the more conventional laboratory methods of virus culture and rapid direct antigen detection tests (7,12,20,22,34). Both laboratory-developed and commercial molecular assays are now available, but the overall complexity of most of these tests has made implementation challenging for many clinical laboratories (18). However, significant recent advances in microfluidics, microelectronics, and microfabrication have allowed the development of simplified molecular systems t...

Evaluation of Simplexa Flu A/B & RSV for Direct Detection of Influenza Viruses (A and B) and Respiratory Syncytial Virus in Patient Clinical Samples

“…Of importance was the RNase P internal control of the LDA, which gave a positive signal for all the samples tested, while the internal control of the Simplexa Flu A/B & RSV kit failed in 4 patient samples (2.3%) that were negative for influenza viruses and RSV. The Simplexa Flu A/B & RSV kit failure rate was higher than the 1.5% reported for other fully automated assays, such as the eSensor respiratory viral panel (12). Upon stratifying the patient samples by virus type, the LDA detected 30 influenza A viruses, whereas the Simplexa Flu A/B & RSV kit detected 28.…”

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.…”

The increasing application of multiplex nucleic acid detection tests to the diagnosis of syndromic infections

“…This announcement reflects the current move towards the development and commercialization of detection technologies for the diagnosis of infectious syndromes whether, enteric [1][2][3][4][5][6], respiratory [7][8][9][10][11][12][13], sexually transmitted [14], affecting the central nervous system [15] or causing sepsis [16,17]. Commercially available tests for the detection of specific pathogens associated with urinary tract infection, neonatal infection or infection in the immunocompromised host have also been developed by commercial diagnostic companies including Fast-track Diagnostics (Junglinster, Luxembourg), AusDiagnostics Pty Ltd (Sydney, Australia) and Seegene (Seoul, Korea).…”