Improved Subtyping of Avian Influenza Viruses Using an RT-qPCR-Based Low Density Array: ‘Riems Influenza a Typing Array’, Version 2 (RITA-2)

Hassan, Kareem E.; Ahrens, Ann Kathrin; Ali, Ahmed; El‐Kady, Magdy F.; Hafez, Hafez M.; Mettenleiter, Thomas C.; Beer, Martin; Harder, Timm C.

doi:10.3390/v14020415

Cited by 28 publications

(28 citation statements)

References 37 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…A comprehensive literature review was carried out to identify available molecular methods for H9 diagnosis. rRT-PCR protocols fully validated and/or extensively used under field conditions, namely Saito et al (2018) [52] and Hassan et al (2019) [29] (later implemented in [64]), were selected to be compared with the new pan-H9 rRT-PCR and the original protocol by Monne et al (2008) [48]. For this purpose, a subset of 46 H9-positive field samples employed for DSe were tested, applying the experimental conditions as per original papers.…”

Section: Comparison With Existing Molecular Diagnostic Methodsmentioning

confidence: 99%

Redesign and Validation of a Real-Time RT-PCR to Improve Surveillance for Avian Influenza Viruses of the H9 Subtype

Panzarin

Marciano

Fortin

et al. 2022

Viruses

View full text Add to dashboard Cite

Avian influenza viruses of the H9 subtype cause significant losses to poultry production in endemic regions of Asia, Africa and the Middle East and pose a risk to human health. The availability of reliable and updated diagnostic tools for H9 surveillance is thus paramount to ensure the prompt identification of this subtype. The genetic variability of H9 represents a challenge for molecular-based diagnostic methods and was the cause for suboptimal detection and false negatives during routine diagnostic monitoring. Starting from a dataset of sequences related to viruses of different origins and clades (Y439, Y280, G1), a bioinformatics workflow was optimized to extract relevant sequence data preparatory for oligonucleotides design. Analytical and diagnostic performances were assessed according to the OIE standards. To facilitate assay deployment, amplification conditions were optimized with different nucleic extraction systems and amplification kits. Performance of the new real-time RT-PCR was also evaluated in comparison to existing H9-detection methods, highlighting a significant improvement of sensitivity and inclusivity, in particular for G1 viruses. Data obtained suggest that the new assay has the potential to be employed under different settings and geographic areas for a sensitive detection of H9 viruses.

show abstract

Section: Comparison With Existing Molecular Diagnostic Methodsmentioning

confidence: 99%

Redesign and Validation of a Real-Time RT-PCR to Improve Surveillance for Avian Influenza Viruses of the H9 Subtype

Panzarin

Marciano

Fortin

et al. 2022

Viruses

View full text Add to dashboard Cite

show abstract

“…Gross pathologic alterations (fibrinous pericarditis, swollen hyperemic liver, spleen, and kidneys) were indicative of a severe infectious disease, which led to an acute death of the young eagle. We analyzed organ samples for AIV by using quantitative reverse transcription PCR as described ( 13 ). HPAIV of subtype H5N1 was found at high viral loads in all tissue samples examined, including the brain (cycle threshold 16.2).…”

Section: Incursion Of Hpaiv H5n1 Into Icelandmentioning

confidence: 99%

“…Arrows indicate viral genomes during 2021 and 2022 in Iceland and assigned to different hemagglutinin clusters B1 and B2. Method hints and basic data are presented in Hassan et al ( 13 ). Scale bar indicates nucleotide substitutions per site.…”

Section: Phylogeographic Identification Of > 2 Vir...mentioning

confidence: 99%

Iceland as Stepping Stone for Spread of Highly Pathogenic Avian Influenza Virus between Europe and North America

Günther¹,

Krone²,

Svansson³

et al. 2022

Emerg. Infect. Dis.

Self Cite

View full text Add to dashboard Cite

Highly pathogenic avian influenza viruses (HPAIVs) of hemagglutinin type H5 and clade 2.3.4.4b have widely spread within the northern hemisphere since 2020 and threaten wild bird populations, as well as poultry production. We present phylogeographic evidence that Iceland has been used as a stepping stone for HPAIV translocation from northern Europe to North America by infected but mobile wild birds. At least 2 independent incursions of HPAIV H5N1 clade 2.3.4.4b assigned to 2 hemagglutinin clusters, B1 and B2, are documented for summer‒autumn 2021 and spring 2022. Spread of HPAIV H5N1 to and among colony-breeding pelagic avian species in Iceland is ongoing. Potentially devastating effects (i.e., local losses >25%) on these species caused by extended HPAIV circulation in space and time are being observed at several affected breeding sites throughout the North Atlantic.

show abstract

“…Recently, a PCR-based panel assay for the detection of multiple poultry respiratory pathogens has been established using a real-time PCR system in a single reaction with SYBR Green reagent ( 3 ). A mini version of the Riems Influenza a Typing Array (RITA) assay ( 34 , 35 ) has also been used in the subtype detection of AIVs and identification of other avian viral pathogens in Bangladesh ( 20 , 36 ). The RITA assay is very useful for detecting the AIV subtype in a single run, but it is not cost-effective for a country with limited resources.…”

Section: Introductionmentioning

confidence: 99%

“…The aim of the study was to detect respiratory viral pathogens (AIV, NDV, IBV, and ILT) and selective subtyping of AIVs using two multitarget real-time quantitative polymerase chain reaction (RT-qPCR) simultaneous detection assays from outbreak samples. We adopt previously established assays ( 34 , 35 ) in a format of multitarget simultaneous runs for the detection of respiratory viral pathogens that are common and endemic in Bangladesh.…”

Section: Introductionmentioning

confidence: 99%

Investigation of respiratory disease outbreaks of poultry in Bangladesh using two real-time PCR-based simultaneous detection assays

et al. 2022

View full text Add to dashboard Cite

For rapid and sensitive pathogen screening from field outbreaks, molecular techniques such as qPCR-based simultaneous detections are efficient. Respiratory diseases are the most detrimental diseases to the poultry industry and need to be addressed because of their major economic losses. In the current study, we have applied two different detection assays: one for simultaneous detection of avian influenza virus (AIV; M gene) and subtyping (H5, N1, H9, N2) using TaqMan probe chemistry (TaqMan multitarget) and another for simultaneous detection of Newcastle disease virus (NDV), infectious bronchitis virus (IBV), and infectious laryngotracheitis virus (ILTV) using SYBR Green chemistry (SYBR Green multitarget). Two individual qPCRs were conducted for the detection of four pathogens. Surveillance of tissue (n = 158) and oropharyngeal swab (206) samples from multiple poultry flocks during the years April 2020–July 2022 applying the TaqMan and SYBR Green multitarget qPCRs revealed that 48.9% of samples were positive for respiratory infections, of which 17.2% were positive for NDV, 25.5% were positive for AIV, 9.9% were positive for IBV, and only a single positive (0.3%) for ILTV. Among the AIV, 35% were highly pathogenic subtype H5N1 and 65% were low pathogenic subtype H9N2. Co-infections of 2–3 respiratory viruses were also accurately detected. Respiratory viral pathogens are quite common in Bangladeshi poultry and can be successfully detected using multitarget simultaneous real-time quantitative polymerase chain reaction (RT-qPCR) assays like those adopted in the current study. Increased mass surveillance, along with the molecular characterization of the circulating respiratory viruses, is crucial to control the epidemic and subsequently save the Bangladeshi poultry industry.

show abstract

Improved Subtyping of Avian Influenza Viruses Using an RT-qPCR-Based Low Density Array: ‘Riems Influenza a Typing Array’, Version 2 (RITA-2)

Cited by 28 publications

References 37 publications

Redesign and Validation of a Real-Time RT-PCR to Improve Surveillance for Avian Influenza Viruses of the H9 Subtype

Redesign and Validation of a Real-Time RT-PCR to Improve Surveillance for Avian Influenza Viruses of the H9 Subtype

Iceland as Stepping Stone for Spread of Highly Pathogenic Avian Influenza Virus between Europe and North America

Investigation of respiratory disease outbreaks of poultry in Bangladesh using two real-time PCR-based simultaneous detection assays

Contact Info

Product

Resources

About