Evidence map›Paper›PMID 41045234›Full record

ArticleJournal of veterinary science2025

Development of a multiplex and universal RT-PCR assay for hemagglutinin and neuraminidase subtyping of avian influenza virus.

Se-Hee An, Gyeong-Beom Heo, Yong-Myung Kang, Youn-Jeong Lee, Kwang-Nyeong Lee

Abstract read
PubMed Full text at PMC Publisher
In one paragraph

Article in Journal of veterinary science, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 1 paper.

0numbers the graph read from it
0cells of the map it votes in
1citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

1 citing paper in PubMed.

  1. Review
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

5 authors.

Se-Hee AnAvian Influenza Research & Diagnostic Division, Animal and Plant Quarantine Agency, Gimcheon 39660, Korea.ORCID https://orcid.org/0000-0002-2206-1299
Gyeong-Beom HeoAvian Influenza Research & Diagnostic Division, Animal and Plant Quarantine Agency, Gimcheon 39660, Korea.ORCID https://orcid.org/0000-0001-8422-8237
Yong-Myung KangCollege of Veterinary Medicine & Institute for Veterinary Biomedical Science, Kyungpook National University, Daegu 41566, Korea.ORCID https://orcid.org/0000-0002-0353-9138
Youn-Jeong LeeAvian Influenza Research & Diagnostic Division, Animal and Plant Quarantine Agency, Gimcheon 39660, Korea.ORCID https://orcid.org/0000-0001-6781-2394
Kwang-Nyeong LeeAvian Influenza Research & Diagnostic Division, Animal and Plant Quarantine Agency, Gimcheon 39660, Korea. leekwn@korea.kr.ORCID https://orcid.org/0000-0002-8267-3983

Funding

Animal and Plant Quarantine Agency B-1543418-2022-23-02
6 · The paper itself

Abstract

importanceAvian influenza virus (AIV) is classified into subtypes by hemagglutinin (HA) and neuraminidase (NA) genes. Wild waterfowl harbor H1-H16 and N1-N9 subtypes. Large-scale AIV surveillance requires substantial labor, time, and cost when using conventional HA and NA subtyping methods.

objectiveThis study aimed to develop multiplex reverse transcription polymerase chain reaction (RT-PCR) and universal assays to detect all H1-H15 and N1-N9 subtypes in just eight RT-PCR reactions.

methodsSubtype-specific forward primers and conserved reverse primers were used to identify HA and NA subtypes based on RT-PCR amplicon size. The HA cleavage site was characterized by sequencing. For NA, a universal forward primer enabled sequencing-based subtyping from a single reaction with the universal reverse primer.

resultsSeventy reference strains and 113 low pathogenic AIVs (from 2021-2022 and 2022-2023 season), covering H1-H15 and N1-N9 subtypes, were tested. The performance of this method was nearly complete, with a few subtype/cleavage site exceptions in relatively long-stored samples. The assays showed 98.90% sensitivity for HA (181/183), 98.91% for NA (182/184), and 100% specificity for both of HA and NA subtyping. CONCLUSIONS AND RELEVANCE: These subtyping assays offer a rapid, sensitive, and cost-effective method for AIV subtyping, enhancing efficiency in large-scale surveillance.

Indexed as

Hemagglutinin Glycoproteins, Influenza VirusInfluenza A virusInfluenza in BirdsMultiplex Polymerase Chain ReactionNeuraminidaseReverse Transcriptase Polymerase Chain ReactionAnimalsBirdsSensitivity and SpecificityHemagglutinin Glycoproteins, Influenza VirusNeuraminidasediagnosisHA and NA subtypeMultiplex PCRorthomyxoviridaeuniversal PCR

Identifiers

PMID41045234
PMCPMC12511891

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.