Impact of Microarray Technology in Influenza Virus Research and Diagnostics

Mukherjee, Sanjay

doi:10.4172/jpb.s6-002

Cited by 2 publications

(3 citation statements)

References 43 publications

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“…These assays typically rely on a single amplified target as an indicator of the presence of a pathogen. While numerous microarray-based assays utilizing multiple probes per target for the detection of human and animal-origin influenza have appeared (Mukherjee and Chakrabarti, 2012;Heydarov et al, 2017;Paulin et al, 2014;Shi et al, 2014;Ryabinin et al, 2011;Gall et al, 2009;Li et al, 2009), none are broadly commercially available. Next generation sequencing of influenza viruses provides the ability for complete characterization and is now routine in the three National Influenza Surveillance Reference Centers and at the CDC.…”

Section: Introductionmentioning

confidence: 99%

Analytical evaluation of the microarray-based FluChip-8G Influenza A+B Assay

Taylor

Dawson

Blair

et al. 2019

Journal of Virological Methods

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A B S T R A C TBackground: Influenza causes a significant annual disease burden, with characterization of the infecting virus important in clinical and public health settings. Rapid immunoassays are fast but insensitive, whereas real-time RT-PCR is sensitive but susceptible to genetic mutations and often requires multiple serial assays. The FluChip-8G Influenza A+B Assay provides type and subtype/lineage identification of influenza A and B, including nonseasonal A viruses, in a single microarray-based assay with same day turnaround time. Objective: To evaluate key analytical performance characteristics of the FluChip-8G Influenza A+B Assay. Study design: Analytical sensitivity, cross-reactivity, and multi-site reproducibility were evaluated. Results: The limit of detection (LOD) for the FluChip-8G influenza A+B Assay ranged from 5.8 × 10 2 -1.5 × 10 5 genome copies/mL, with most samples ∼2 × 10 3 genome copies/mL (∼160 genome copies/reaction). Fifty two (52) additional strains were correctly identified near the LOD, demonstrating robust reactivity. Two variant viruses (H1N1v and H3N2v) resulted in dual identification as both "non-seasonal influenza A" and A/ H1N1pdm09. No reproducible cross-reactivity was observed for the 34 organisms tested, however, challenges with internal control inhibition due to crude growth matrix were observed. Lastly, samples tested near the LOD showed high reproducibility (97.0% (95% CI 94.7-98.7)) regardless of operator, site, reagent lot, or testing day. Conclusion: The FluChip-8G Influenza A+B Assay is an effective new method for detecting and identifying both seasonal and non-seasonal influenza viruses, as revealed by good sensitivity and robust reactivity to 52 unique strains of influenza virus. In addition, the lack of cross-reactivity to non-influenza pathogens and high lab-to-lab reproducibility highlight the analytical performance of the assay as an alternative to real-time RT-PCR and sequencing-based assays. Clinical validation of the technology in a multi-site clinical study is the subject of a separate investigation.

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Section: Introductionmentioning

confidence: 99%

Analytical evaluation of the microarray-based FluChip-8G Influenza A+B Assay

Taylor

Dawson

Blair

et al. 2019

Journal of Virological Methods

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“…DNA array also known as DNA chip is a collection of spots attached to a solid support where each spot can contain one or more probes. Microarray is simply the arrangement of biomolecules on a solid surface in order to generate qualitative and quantitative information [55,56]. It is a technology used to identify LASU Journal of Research and Review in Science labelled targets by hybridizing them with specific probes and hybridization signals are mapped within the array.…”

Section: Dna Microarraymentioning

confidence: 99%

“…It has a high throughput because thousands of probes can be spotted on a single slide [56]. It can detect thousands of targets at the same time and can be used to prove phylogenetic relationships between isolates [58].…”

Section: Dna Microarraymentioning

confidence: 99%

Understanding Nucleic Acid Amplification Techniques in the Detection of Influenza viruses in Developing Countries

Anjorin

Salu

Robert

et al. 2019

JRRS

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Introduction: Early detection of emerging influenza virus variant is a key factor in the WHO influenza Global strategies for prevention and control. Rapid, accurate, inexpensive and portable detection systems are needed for influenza virus diagnosis and surveillance. Such a detection system should easily identify all the subtypes of influenza virus. Degenerate primers and probes designed from evolutionally conserved regions for known influenza A viruses present the best way to identify unknown subtypes of influenza A virus by polymerase chain reaction PCR and array techniques. The isothermal reactions, Nucleic Acid Sequencing Based Amplification (NASBA) and Loop-mediated isothermal Amplification (LAMP) possess great potential for influenza A virus detection especially in developing countries. However, multiplex real-time (rT) or quantitative (q) polymerase chain reaction (qPCR) remains a rapid, accurate and timesaving technique used for influenza virus detection. Aim: This manuscript explained the principles of nucleic acid amplification techniques commonly used in developing countries. Methods: Literature search was done in NCBI PUBMED, PUBMED Central and Google Scholar using words and phrases including “Influenzamolecular diagnosis, NAAT”, Molecular techniques/ methods, PCR, qPCR, NASBA, LAMP, and DNA microarray. Results: The underlining principles and basic processes involved in the application of nucleic acid amplification techniques for the detection and epidemiological surveillance of influenza virus were identified and grouped under PCR (RT-PCR and qRT-PCR) and Non-PCR (LCR, pyrosequencing, NASBA, LAMP and DNA microarray) amplifications. Conclusion: It is hoped that by understanding the techniques and basic principles of Nucleic acid amplifications, less expensive, and more convenient protocols for influenza virus detection and surveillance can be developed Keywords: Influenza, NAAT, Molecular, PCR, qPCR, Viral diagnosis.

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Impact of Microarray Technology in Influenza Virus Research and Diagnostics

Cited by 2 publications

References 43 publications

Analytical evaluation of the microarray-based FluChip-8G Influenza A+B Assay

Analytical evaluation of the microarray-based FluChip-8G Influenza A+B Assay

Understanding Nucleic Acid Amplification Techniques in the Detection of Influenza viruses in Developing Countries

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