scholarly journals The Panther Fusion System with Open Access Functionality for Laboratory-Developed Tests for Influenza A Virus Subtyping

2020 ◽  
Vol 58 (6) ◽  
Author(s):  
Kathleen A. Stellrecht ◽  
Jesse L. Cimino ◽  
Vincente P. Maceira
Keyword(s):  
Open Access ◽  
Influenza A ◽  
Limit Of Detection ◽  
Turnaround Time ◽  
Nucleic Acid Amplification ◽  
Influenza B ◽  
Fusion System ◽  
Valuable Complement ◽  
Syncytial Virus

ABSTRACT Nucleic acid amplification tests, such as PCR, are the method of choice for respiratory virus testing, due to their superior diagnostic accuracy and fast turnaround time. The Panther Fusion (Fusion; Hologic) system has an array of highly sensitive in vitro diagnostic (IVD) real-time PCR assays for respiratory viruses, including an assay for influenza A (FluA) virus, influenza B (FluB) virus, and respiratory syncytial virus (RSV) (FFABR assay). The Fusion system has Open Access functionality to perform laboratory-developed tests (LDTs) alongside IVD assays. We developed two LDTs for FluA virus strain typing on the Panther Fusion instrument, enabling side-by-side testing with the FFABR assay. The LDT-FAST assay uses proprietary primers and probes designed by Hologic for the Prodesse ProFAST+ (PFAST) assay. The exWHO-FAST assay is an expanded redesign of the WHO-recommended reverse transcriptase PCRs (RT-PCRs). To evaluate the performance of these two LDTs, 110 FluA virus-positive samples were tested. Of these, 104 had been subtyped previously; 54 were H3, 46 were 09H1, and 4 were fsH1. All were appropriately subtyped by both LDTs. Of the untyped FluA virus samples, three were subtyped as H3 by both LDTs and two were subtyped as H3 by the LDT-FAST assay only. The sample not subtyped by either LDT was retested with the FFABR assay and was now negative. Limit-of-detection (LOD) analyses were performed with five FluA virus strains. The LDT-FAST LODs were similar to the FFABR assay LODs, while the exWHO-FAST LODs were higher for two H3N2 strains, findings that were explained by analysis of primer/probe homology. In conclusion, either FluA virus typing assay would be a valuable complement to the Panther Fusion respiratory menu given the performance of these LDTs, the system’s full automation, and the ability to split eluates for both IVD and LDT testing.

scholarly journals Clinical and Financial Benefits of Rapid Detection of Respiratory Viruses: an Outcomes Study

2000 ◽  
Vol 38 (8) ◽  
pp. 2824-2828 ◽  
Author(s):  
Joan Barenfanger ◽  
Cheryl Drake ◽  
Nidia Leon ◽  
Tina Mueller ◽  
Tammy Troutt
Keyword(s):  
Length Of Stay ◽  
Influenza A ◽  
Respiratory Viruses ◽  
Turnaround Time ◽  
Influenza B Virus ◽  
Influenza B ◽  
Shell Vial ◽  
The Mean ◽  
Syncytial Virus ◽  
Standard Techniques

To assess the expected benefits of rapid reporting of respiratory viruses, we compared patients whose samples were processed using standard techniques such as enzyme immunoassays, shell vial assays, and culture tube assays (year 1) to patients whose samples were processed with the same standard techniques in addition to immunofluorescent testing (FA) directly on cytocentrifuged samples (year 2). The cytospin FA screened for influenza A and B viruses, respiratory syncytial virus (RSV), parainfluenza viruses 1 to 3, and adenovirus (DAKO Diagnostics Ltd.). The specificity of the cytospin FA for all viruses was 100%. The sensitivities for influenza A virus and RSV were 90 and 98%, respectively, but the sensitivities for influenza B virus and adenovirus were unacceptable (14.3 and 0%, respectively). However, since the former viruses account for >85% of our isolates from clinical specimens, the cytospin FA is an excellent screening test since the positive result was available within hours. The mean turnaround time for all positive viruses was 4.5 days in year 1 and 0.9 day in year 2 (P = 0.001). This rapid reporting resulted in physicians having access to information sooner, enabling more appropriate treatment. The mean length of stay in the hospital for inpatients with respiratory viral isolates was 10.6 days for year 1 versus 5.3 days for year 2. Mean variable costs for these patients was $7,893 in year 1 and $2,177 in year 2. After subtracting reagent costs and technological time, the savings in variable costs was $144,332/year. Summarizing, the cytospin FA markedly decreased turnaround time and was associated with decreased mortality, length of stay, and costs and with better antibiotic stewardship.

scholarly journals Multicenter Evaluation of the QIAstat-Dx Respiratory Panel for Detection of Viruses and Bacteria in Nasopharyngeal Swab Specimens

2020 ◽  
Vol 58 (5) ◽  
Author(s):  
Amy L. Leber ◽  
Jan Gorm Lisby ◽  
Glen Hansen ◽  
Ryan F. Relich ◽  
Uffe Vest Schneider ◽  
...  
Keyword(s):  
Influenza A ◽  
Parainfluenza Virus ◽  
Nasopharyngeal Swab ◽  
Influenza B ◽  
Respiratory Pathogens ◽  
Content Type ◽  
Percent Agreement ◽  
Influenza A H1n1 ◽  
Syncytial Virus

ABSTRACT The QIAstat-Dx Respiratory Panel (QIAstat-Dx RP) is a multiplex in vitro diagnostic test for the qualitative detection of 20 pathogens directly from nasopharyngeal swab (NPS) specimens. The assay is performed using a simple sample-to-answer platform with results available in approximately 69 min. The pathogens identified are adenovirus, coronavirus 229E, coronavirus HKU1, coronavirus NL63, coronavirus OC43, human metapneumovirus A and B, influenza A, influenza A H1, influenza A H3, influenza A H1N1/2009, influenza B, parainfluenza virus 1, parainfluenza virus 2, parainfluenza virus 3, parainfluenza virus 4, rhinovirus/enterovirus, respiratory syncytial virus A and B, Bordetella pertussis, Chlamydophila pneumoniae, and Mycoplasma pneumoniae. This multicenter evaluation provides data obtained from 1,994 prospectively collected and 310 retrospectively collected (archived) NPS specimens with performance compared to that of the BioFire FilmArray Respiratory Panel, version 1.7. The overall percent agreement between QIAstat-Dx RP and the comparator testing was 99.5%. In the prospective cohort, the QIAstat-Dx RP demonstrated a positive percent agreement of 94.0% or greater for the detection of all but four analytes: coronaviruses 229E, NL63, and OC43 and rhinovirus/enterovirus. The test also demonstrated a negative percent agreement of ≥97.9% for all analytes. The QIAstat-Dx RP is a robust and accurate assay for rapid, comprehensive testing for respiratory pathogens.

scholarly journals Direct Comparison of Alere i and cobas Liat Influenza A and B Tests for Rapid Detection of Influenza Virus Infection

2016 ◽  
Vol 54 (11) ◽  
pp. 2763-2766 ◽  
Author(s):  
Frederick S. Nolte ◽  
Lori Gauld ◽  
Susan B. Barrett
Keyword(s):  
Influenza A Virus ◽  
Influenza A ◽  
Limit Of Detection ◽  
Point Of Care ◽  
Influenza Virus Infection ◽  
Nucleic Acid Amplification ◽  
Influenza B Virus ◽  
Influenza B ◽  
Viral Transport Medium ◽  
B Virus

We compared two rapid, point-of care nucleic acid amplification tests for detection of influenza A and B viruses (Alere i [Alere] and cobas Liat [Roche Diagnostics]) with the influenza A and B virus test components of the FilmArray respiratory panel (BioFire Diagnostics) using 129 respiratory specimens collected in universal viral transport medium (80 influenza A virus and 16 influenza B virus positive) from both adult and pediatric patients. The sensitivities of the Alere test were 71.3% for influenza A virus and 93.3% for influenza B virus, with specificities of 100% for both viruses. The sensitivities and specificities of the Liat test were 100% for both influenza A and B viruses. The poor sensitivity of the Alere test for detection of influenza A virus was likely due to a study set that included many low-positive samples that were below its limit of detection.

scholarly journals Comparison of Six Sample-to-Answer Influenza A/B and Respiratory Syncytial Virus Nucleic Acid Amplification Assays Using Respiratory Specimens from Children

2018 ◽  
Vol 56 (11) ◽  
Author(s):  
Dithi Banerjee ◽  
Neena Kanwar ◽  
Ferdaus Hassan ◽  
Cynthia Essmyer ◽  
Rangaraj Selvarangan
Keyword(s):  
Respiratory Syncytial Virus ◽  
Nucleic Acid ◽  
Influenza A ◽  
Nucleic Acid Amplification ◽  
Influenza B Virus ◽  
Influenza B ◽  
Reverse Transcription Pcr ◽  
B Virus ◽  
Control And Prevention ◽  
Syncytial Virus

ABSTRACT The rapid and accurate detection of influenza A virus (FluA), influenza B virus (FluB), and respiratory syncytial virus (RSV) improves patient care. Sample-to-answer (STA) platforms based on nucleic acid amplification and detection of these viruses are simple, automated, and accurate. We compared six such platforms for the detection of FluA, FluB, and RSV: Cepheid GeneXpert Xpress Flu/RSV (Xpert), Hologic Panther Fusion Flu A/B/RSV (Fusion), Cobas influenza A/B & RSV (Liat), Luminex Aries Flu A/B & RSV (Aries), BioFire FilmArray respiratory panel (RP), and Diasorin Simplexa Flu A/B & RSV (Simplexa). Nasopharyngeal (NP) swab specimens (n = 225) from children previously tested by RP were assessed on these platforms. The results were compared to those of the Centers for Disease Control and Prevention (CDC)-developed real-time reverse transcription-PCR (rRT-PCR) assay for influenza A/B viruses and RSV. Subtyping for FluA and FluB was performed for discrepant analysis where applicable. The percent sensitivities/specificities for FluA detection were 100/100 (Fusion), 98.6/99.3 (Xpert), 100/100 (Liat), 98.6/100 (Aries), 98.6/100 (Simplexa), and 100/100 (RP). The percent sensitivities/specificities for FluB detection were 100/100 (Fusion), 97.9/99.4 (Xpert), 97.9/98.3 (Liat), 93.7/99.4 (Aries), 85.4/99.4 (Simplexa), and 95.8/97.7 (RP); and those for RSV detection were 98.1/99.4 (Xpert), 98.1/99.4 (Liat), 96.3/100 (Fusion), 94.4/100 (Aries), 87/94.4 (Simplexa), and 94.4/100 (RP). The 75 strains confirmed to be FluA included 29 pH1N1, 39 H3N2, 4 sH1N1, and 3 untyped strains. The 48 strains confirmed to be FluB included 33 strains of the Yamagata lineage, 13 of the Victoria lineage, 1 of both the Yamagata and Victoria lineages, and 1 of an unknown lineage. All six STA platforms demonstrated >95% sensitivity for FluA detection, while three platforms (Fusion, Xpert, and Liat) demonstrated >95% sensitivity for FluB and RSV detection.

scholarly journals Orally Efficacious Broad-Spectrum Ribonucleoside Analog Inhibitor of Influenza and Respiratory Syncytial Viruses

2018 ◽  
Vol 62 (8) ◽  
Author(s):  
Jeong-Joong Yoon ◽  
Mart Toots ◽  
Sujin Lee ◽  
Myung-Eun Lee ◽  
Barbara Ludeke ◽  
...  
Keyword(s):  
Broad Spectrum ◽  
High Throughput Screening ◽  
Influenza A ◽  
Influenza Viruses ◽  
Transmission Model ◽  
Mouse Lung ◽  
Influenza B ◽  
Good Tolerability ◽  
Syncytial Virus

ABSTRACT Morbidity and mortality resulting from influenza-like disease are a threat, especially for older adults. To improve case management, next-generation broad-spectrum antiviral therapeutics that are efficacious against major drivers of influenza-like disease, including influenza viruses and respiratory syncytial virus (RSV), are urgently needed. Using a dual-pathogen high-throughput screening protocol for influenza A virus (IAV) and RSV inhibitors, we have identified N4-hydroxycytidine (NHC) as a potent inhibitor of RSV, influenza B viruses, and IAVs of human, avian, and swine origins. Biochemical in vitro polymerase assays and viral RNA sequencing revealed that the ribonucleotide analog is incorporated into nascent viral RNAs in place of cytidine, increasing the frequency of viral mutagenesis. Viral passaging in cell culture in the presence of an inhibitor did not induce robust resistance. Pharmacokinetic profiling demonstrated dose-dependent oral bioavailability of 36 to 56%, sustained levels of the active 5′-triphosphate anabolite in primary human airway cells and mouse lung tissue, and good tolerability after extended dosing at 800 mg/kg of body weight/day. The compound was orally efficacious against RSV and both seasonal and highly pathogenic avian IAVs in mouse models, reducing lung virus loads and alleviating disease biomarkers. Oral dosing reduced IAV burdens in a guinea pig transmission model and suppressed virus spread to uninfected contact animals through direct transmission. Based on its broad-spectrum efficacy and pharmacokinetic properties, NHC is a promising candidate for future clinical development as a treatment option for influenza-like diseases.

scholarly journals Inhibition of Respiratory Syncytial Virus Replication in vitro by a Pyrazole Dicarboxamide Analogue of Ribavirin

1994 ◽  
Vol 5 (5) ◽  
pp. 340-343 ◽  
Author(s):  
J. M. Woods ◽  
C. L. P. Marr ◽  
C. R. Penn
Keyword(s):  
Respiratory Syncytial Virus ◽  
Virus Replication ◽  
Influenza A ◽  
Adenosine Kinase ◽  
Influenza B ◽  
Inosine Monophosphate Dehydrogenase ◽  
Inosine Monophosphate ◽  
Syncytial Virus ◽  
Monophosphate Dehydrogenase

1-β-D-Ribofuranosylpyrazole-3,4-dicarboxamide (GR-92938X) was found to be an inhibitor of respiratory syncytial virus in vitro, with a potency equivalent to that of ribavirin. The compound was more specific as, unlike ribavirin, it did not inhibit influenza A, influenza B or parainfluenza 2 virus. It was phosphorylated by adenosine kinase and, unlike ribavirin, it did not inhibit cellular inosine monophosphate dehydrogenase.

scholarly journals Verification of a Novel Multiplex PCR Respiratory Virus Panel in a US Biocontainment Unit

2019 ◽  
Vol 152 (Supplement_1) ◽  
pp. S7-S7
Author(s):  
Christina L Dean ◽  
Emily Alvey ◽  
Crystal Evans ◽  
Charles Hill ◽  
Eileen Burd ◽  
...  
Keyword(s):  
Influenza A ◽  
Respiratory Virus ◽  
Limit Of Detection ◽  
Communicable Disease ◽  
Control Sample ◽  
The United States ◽  
Clinical Samples ◽  
Influenza B ◽  
Respiratory Pathogens ◽  
Syncytial Virus

Abstract Emerging infectious diseases carry unique logistical, financial, and clinical ramifications. Rapid diagnostic testing methods can alleviate some of these challenges by providing definitive diagnoses earlier in the clinical course, leading to appropriate targeted therapy, cost savings, and improved patient outcomes. The BioFire FilmArray Respiratory Panel 2 plus (RP2plus; bioMérieux, Marcy l’Etoile, France) is a multiplexed nucleic acid test for detection of Middle East respiratory syndrome coronavirus (MERS-CoV) and 14 common viral and 4 bacterial respiratory pathogens in nasopharyngeal swabs obtained from those meeting MERS-CoV epidemiological criteria. The aim of this study was to verify the FilmArray RP2plus for use in our biocontainment unit. Of note, the RP2plus is FDA approved but not currently available for sale in the United States. Eight patient samples were tested with known results (GenMark Respiratory Virus Panel [RVP] or Cepheid Xpert Flu/RSV). We had concordant results between the platforms for samples containing influenza A, respiratory syncytial virus (RSV), parainfluenza virus 2, rhinovirus, and a negative sample. We evaluated two influenza B samples from two different patients. The FilmArray RP2plus did not detect influenza B in one of the patient samples. The sample was exhausted and repeat testing could not be performed. A second rhinovirus sample was not detected by the RP2plus, but Coronavirus 229E was detected in this sample, a virus not detected by the RVP. The sample was repeated and again did not detect rhinovirus. Further investigation into this discrepancy revealed that rhinovirus was originally detected by RVP at a signal of 34.4 nA (repeat of 46.9 nA). The concordant rhinovirus sample had a signal of 226.7 nA by RVP, which was much higher than the discrepant sample. Because of the low signal by RVP in the discrepant sample, perhaps the viral load was below the limit of detection of the RP2plus. All other quality control sample pools passed verification testing, including day-to-day and operator variance. It is not uncommon for a person under investigation (PUI) for a highly communicable disease to be evaluated in our facility. The performance of the RP2plus test on clinical samples showed acceptable concordance with our current means of testing for respiratory pathogens. The RP2plus will eliminate challenges implicated in storing and transporting specimens to an off-site lab, facilitate quicker turnaround time, and streamline the often cumbersome, complex protocols and practices required to work up a serious communicable disease.

scholarly journals Kinetics Of Interferon-λ And Receptor Expression In Response To In Vitro Respiratory Viral Infection

2021 ◽  
Author(s):  
Alexey Lozhkov ◽  
Nikita Yolshin ◽  
Irina Baranovskaya ◽  
Marina Plotnikova ◽  
Mariia Sergeeva ◽  
...  
Keyword(s):  
Influenza A ◽  
Alveolar Epithelial Cell ◽  
Mrna Level ◽  
Receptor Expression ◽  
Influenza B Virus ◽  
Influenza B ◽  
Type I ◽  
Viral Spread ◽  
Syncytial Virus

The major protective immune response against viruses is production of type I and III interferons (IFNs). IFNs induce the expression of hundreds of IFN-stimulated genes (ISGs) that block viral replication and further viral spread. The ability of respiratory viruses to suppress induction of IFN-mediated antiviral defenses in infected epithelial cells may be a factor contributing to the particular pathogenicity of several strains. In this report, we analyzed expression of IFNs and some ISGs in an alveolar epithelial cell subtype (A549) in response to infection with: influenza A viruses (A/California/07/09pdm (H1N1), A/Texas/50/12 (H3N2)); influenza B virus (B/Phuket/3073/13); adenovirus type 5 and 6; or respiratory syncytial virus (strain A2). IFNL and ISGs expression significantly increased in response to infection with all RNA viruses 24 hpi. Nevertheless, only IBV led to early increase in IFNL and ISGs mRNA level. IBV and H1N1 infection led to elevated proinflammatory cytokine production. We speculate that augmented IFN-α, IFN-β, IL-6 levels negatively correlate to SOCS1 expression. Importantly, we showed a decrease in IFNLR1 mRNA in case of IBV infection that implies the existence of negative ISGs expression regulation at IFNλR level. It could be either a specific feature of IBV or a consequence of early IFNL expression.

scholarly journals Multi-center Evaluation of the Cepheid Xpert® Xpress SARS-CoV-2/Flu/RSV Test

2020 ◽  
Author(s):  
Heba H. Mostafa ◽  
Karen C. Carroll ◽  
Rachel Hicken ◽  
Gregory J. Berry ◽  
Ryhana Manji ◽  
...  
Keyword(s):  
Public Health ◽  
Los Angeles ◽  
Influenza A ◽  
Respiratory Virus ◽  
Medical Center ◽  
Standard Of Care ◽  
Nucleic Acid Amplification ◽  
Nasopharyngeal Swab ◽  
Influenza B ◽  
Syncytial Virus

With the approach of respiratory virus season in the Northern Hemisphere, clinical microbiology and public health laboratories will need rapid diagnostic assays to distinguish SARS-CoV-2 from influenza and respiratory syncytial virus (RSV) infections for diagnosis and surveillance. In this study, the clinical performance of the Xpert® Xpress SARS-CoV-2/Flu/RSV test (Cepheid, Sunnyvale, CA, USA) for nasopharyngeal swab specimens was evaluated in four centers: Johns Hopkins Medical Microbiology Laboratory, Northwell Health Laboratories, NYC Public Health Laboratory, and Los Angeles County/University of Southern California (LAC+USC) Medical Center. A total of 319 nasopharyngeal swab specimens, positive for SARS-CoV-2 (n = 75), influenza A (n = 65), influenza B (n = 50), RSV (n = 38), or negative (n = 91) by the standard of care nucleic acid amplification tests at each site were tested using the Cepheid panel test. The overall positive percent agreement for the SARS-CoV-2 target was 98.7% (n= 74/75) and the negative agreement was 100% (n= 91) with all other analytes showing 100% total agreement (n= 153). Standard of care tests to which the Cepheid panel was compared included the Cepheid Xpert Xpress SARS-CoV-2, Cepheid Xpert Xpress Flu/RSV, the GenMark ePlex respiratory panel, the BioFire Respiratory panels 2.1 and v1.7, the DiaSorin Simplexa COVID-19 Direct, and the Hologic Panther Fusion SARS-CoV-2 assays. The Xpert Xpress SARS-CoV-2/Flu/RSV test showed high sensitivity and accuracy for all analytes included in the test. This test will provide a valuable clinical diagnostic and public health solution for detecting and differentiating SARS-CoV-2, influenza A and B, and RSV infections during the current respiratory virus season.

Pneumocystis carinii Pneumonitis in Young Immunocompetent Infants

PEDIATRICS ◽  
1980 ◽  
Vol 66 (1) ◽  
pp. 56-62
Author(s):  
Sergio Stagno ◽  
Linda L. Pifer ◽  
Walter T. Hughes ◽  
Dana M. Brasfield ◽  
Ralph E. Tiller
Keyword(s):  
Influenza A ◽  
Pneumocystis Carinii ◽  
Open Lung Biopsy ◽  
Influenza B ◽  
Severe Respiratory Distress ◽  
Number Of Patients ◽  
Circulating Antigens ◽  
Cytomegalovirus Pneumonia ◽  
Syncytial Virus ◽  
A Prospective Study

Of 67 infants enrolled in a prospective study of infant pneumonia ten (14%) had evidence of Pneumocystis carinii infection. Diagnosis was achieved by demonstrating circulating P carinii antigens by counterimmunoelectrophoresis in all ten cases and by histopathology in the only infant who underwent an open lung biopsy. Antigenemia did not occur in 64 control infants (P = .003), nor in 57 patients of similar age who were hospitalized with pneumonitis due to Chlamydia trachomatis, respiratory syncytial virus, cytomegalovirus, adenovirus, and influenza A and influenza B viruses. None of the ten infants with P carinii pneumonitis had evidence of a primary immunodeficiency nor had any received immunosuppressive medication. These patients were hospitalized at a mean age of 6 weeks (range 2 to 12) and their illness was characterized by its afebrile course, presentation in crisis with severe respiratory distress, apnea, tachypnea, cough, increased IgM, and bilateral pulmonary infiltrates with hyperaeration. The clinical features of P carinii pneumonitis were indistinguishable from those of C trachomatis and cytomegalovirus pneumonia. Treatment with trimethoprim-sulfamethoxazole was associated with rapid disappearance of circulating antigens; however, the small number of patients studied did not permit an analysis of its clinical efficacy. These results indicate that P carinii singly or in combination with other infectious agents may be an important cause of pneumonitis in young, immunocompetent infants with no underlying illnesses.

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