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

Hindiyeh, Musa; Kolet, Liat; Meningher, Tal; Weil, Merav; Mendelson, Ella; Mandelboim, Michal

doi:10.1128/jcm.00286-13

Cited by 28 publications

(18 citation statements)

References 13 publications

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“…Several papers were published on Simplexa™ Flu A/B & RSV, using an extracted nucleic acid and/or direct sample testing and comparing to conventional methods or molecular tests (Alby et al, 2013;Hindiyeh et al, 2013;Selvaraju et al, 2014;Ko et al, 2013;Woodberry MW1 et al, 2013;Landry and Ferguson, 2014;Svensson et al, 2014). In this study, the Simplexa™ Flu A/B & RSV Direct assay system showed comparable performance characteristics for Flu A, Flu B and RSV in comparison with TAC testing.…”

Section: Discussionmentioning

confidence: 71%

Performance evaluation of direct fluorescent antibody, Focus Diagnostics Simplexa™ Flu A/B & RSV and multi-parameter customized respiratory Taqman® array card in immunocompromised patients

Steensels

Descheemaeker

Curran

et al. 2017

Journal of Virological Methods

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Section: Discussionmentioning

confidence: 71%

Performance evaluation of direct fluorescent antibody, Focus Diagnostics Simplexa™ Flu A/B & RSV and multi-parameter customized respiratory Taqman® array card in immunocompromised patients

Steensels

Descheemaeker

Curran

et al. 2017

Journal of Virological Methods

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“…Not surprisingly, the results varied. Two studies using fresh samples found Simplexa direct testing (2 and 50 l) to be similar in sensitivity to their LDT PCRs (8,9). The proportion of samples with low viral loads was not reported in those studies.…”

mentioning

confidence: 99%

“…Several papers were recently published on Simplexa Flu A/B & RSV using one or more of three approaches, extracted nucleic acid (5-7) and/or direct testing of either 2 l of sample in a 96-well disc (6,8) or 50 l of sample in an 8-well disc (7,9). The 50-l option is now available as an in vitro diagnostic (IVD) kit.…”

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confidence: 99%

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Comparison of Simplexa Flu A/B & RSV PCR with Cytospin-Immunofluorescence and Laboratory-Developed TaqMan PCR in Predominantly Adult Hospitalized Patients

Landry

Ferguson

2014

J Clin Microbiol

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To compare Simplexa Flu A/B & RSV PCR with cytospin-immunofluorescence and laboratory-developed TaqMan PCR methods, 445 nasopharyngeal samples were tested. Of these, 199 were positive (46 for respiratory syncytial virus [RSV], 120 for influenza A, and 33 for influenza B) and 246 were negative. The direct fluorescent-antibody assay (DFA) detected 132 (66.3%) positive samples, Simplexa direct detected 162 (81.4%), Simplexa using extracts detected 177 (88.9%), and lab-developed TaqMan PCR reference methods detected 199 (100%). The specificities were 99.6%, 100%, 100%, and 100%, respectively. The two Simplexa methods were more sensitive than the DFA (P ‫؍‬ 0.0001) but less sensitive than the TaqMan reverse transcriptase PCR (P ‫؍‬ 0.0001). Influenza virus and respiratory syncytial virus (RSV) cause a substantial disease burden in people of all ages. A rapid laboratory diagnosis can facilitate early antiviral therapy, discontinuation of antibiotics, and implementation of infection control measures (1). Our laboratory uses the cytospin-enhanced direct fluorescent-antibody assay (DFA) as an initial on-demand screen and laboratory-developed real-time TaqMan (LDT) PCR when greater sensitivity is needed (2). In this report, we evaluated Simplexa Flu A/B & RSV PCR as a potential replacement for DFA and LDT PCR.Nasopharyngeal swab specimens submitted in 3 ml M4 medium (Remel, Lenexa, KS) from February to April 2011 were tested by DFA and LDT PCR as requested by the clinician within 2 to 12 h of collection. The original samples and extracts were then frozen at Ϫ70°C, and a subset of the positive and negative samples were retested with the Simplexa PCR within 2 to 4 months.The cytospin-DFA was performed using SimulFluor reagents (Millipore, Billerica, MA) as previously described (3). For influenza detection and subtyping, the CDC real-time reverse transcriptase (RT) PCR primers and probes were used (2). For RSV, the protocol published by van Elden et al. (4) was used, but the subtypes were not differentiated. Total nucleic acid was extracted from 200 l of sample and eluted in 55 l on an EasyMag instrument (bioMérieux, Durham, NC), and 5 l of extract was added to 20 l AgPath-ID one-step RT-PCR master mix (Life Technologies, Carlsbad, CA) without an internal control (IC), and the manufacturers' cycling parameters (10 min at 45°C, 10 min at 95°C, 15 s at 95°C, 45 s at 60°C) were used. Amplification was performed for 45 cycles using an ABI 7500 instrument (Applied Biosystems, Foster City, CA). Positive samples with cycle threshold (C T ) values of Ͻ38 were accepted as positive. For C T values of Ն38, amplification was repeated in duplicate and accepted if one replicate was positive.For Simplexa Flu A/B & RSV PCR (Focus Diagnostics, Cypress, CA), frozen extracts were thawed, vortexed, and pulse spun. To each well of a 96-well disc was added 5 l of master mix containing primers, an IC, and 5 l of thawed extract or positive or negative control material. Discs were sealed and placed in a 3 M integrated cycler (Focus Diagnostics). Amp...

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“…A study comparing LDT to the same Simplexa Flu A/B & RSV assay with a modified protocol that eliminated nucleic acid extraction but used the conventional real-time PCR format reported overall sensitivities of 95.1% and 99.6 (8).…”

mentioning

confidence: 99%

Comparison of the Simplexa FluA/B & RSV Direct Assay and Laboratory-Developed Real-Time PCR Assays for Detection of Respiratory Virus

et al. 2013

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

Cited by 28 publications

References 13 publications

Performance evaluation of direct fluorescent antibody, Focus Diagnostics Simplexa™ Flu A/B & RSV and multi-parameter customized respiratory Taqman® array card in immunocompromised patients

Performance evaluation of direct fluorescent antibody, Focus Diagnostics Simplexa™ Flu A/B & RSV and multi-parameter customized respiratory Taqman® array card in immunocompromised patients

Comparison of Simplexa Flu A/B & RSV PCR with Cytospin-Immunofluorescence and Laboratory-Developed TaqMan PCR in Predominantly Adult Hospitalized Patients

Comparison of the Simplexa FluA/B & RSV Direct Assay and Laboratory-Developed Real-Time PCR Assays for Detection of Respiratory Virus

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