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

被引：11

作者：

Panzarin, Valentina ^{[1
]}

Marciano, Sabrina ^{[1
]}

Fortin, Andrea ^{[1
]}

Brian, Irene ^{[1
]}

D'Amico, Valeria ^{[1
]}

Gobbo, Federica ^{[1
]}

Bonfante, Francesco ^{[1
]}

Palumbo, Elisa ^{[1
]}

Sakoda, Yoshihiro ^{[2
]}

Le, Kien Trung ^{[2
]}

Duc-Huy Chu ^{[3
]}

Shittu, Ismaila ^{[4
]}

Meseko, Clement ^{[4
]}

Haido, Abdoul Malick ^{[5
]}

Odoom, Theophilus ^{[6
]}

Diouf, Mame Nahe ^{[7
]}

Djegui, Fidelia ^{[8
]}

Steensels, Mieke ^{[9
]}

Terregino, Calogero ^{[1
]}

Monne, Isabella ^{[1
]}

机构：

[1] Ist Zooprofilatt Sperimentale Venezie IZSVe, EU OIE Natl Reference Lab Avian Influenza & Newca, FAO Reference Ctr Anim Influenza & Newcastle Dis, Div Comparat Biomed Sci, I-35020 Legnaro, Italy

[2] Hokkaido Univ, OIE Reference Lab Avian Influenza, Fac Vet Med, Sapporo, Hokkaido 0600818, Japan

[3] Minist Agr & Rural Dev MARD, Dept Anim Hlth, Hanoi 11519, Vietnam

[4] Natl Vet Res Inst NVRI, Reg Lab Anim Influenzas & Other Transboundary Ani, Vom 930010, Nigeria

[5] Minist Agr & Elevage, Lab Cent Elevage LABOCEL, Niamey 485, Niger

[6] Minist Food & Agr, Accra Vet Lab, Vet Serv Directorate, M161, Accra, Ghana

[7] Inst Senegalais Rech Agr ISRA, Lab Natl Elevage & Rech Vet LNERV, Dakar 2057, Senegal

[8] Lab Diagnost Vet & Serosurveillance LADISERO, Parakou 23, Benin

[9] Sciensano, AI ND Natl Reference Lab, B-1050 Brussels, Belgium

来源：

VIRUSES-BASEL | 2022年 / 14卷 / 06期

关键词：

avian influenza; H9Nx; molecular diagnosis; real-time RT-PCR; validation; LIVE BIRD MARKETS; MOLECULAR CHARACTERIZATION; SEQUENCE ALIGNMENT; BROILER-CHICKENS; POULTRY; H7; H5; GENE; PREVALENCE; EVOLUTION;

D O I：

10.3390/v14061263

中图分类号：

Q93 [微生物学];

学科分类号：

071005 ; 100705 ;

摘要：

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.

引用

页数：14

共 65 条

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