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Challenges for accurate and prompt molecular diagnosis of clades of highly pathogenic avian influenza H5N1 viruses emerging in Vietnam.
Avian Pathol 2012; 41(2):177-93AP

Abstract

Forty-six chickens and 48 ducks were sampled from four Vietnamese poultry premises in 2009 infected with H5N1 highly pathogenic avian influenza (HPAI) clade 2.3.2 and 2.3.4 viruses, which also differed by cleavage site (CS) sequences in their haemagglutinin (HA) genes. All clinical specimens (n=282), namely tracheal and cloacal swabs plus feathers, were tested by five Eurasian reverse-transcriptase AI RealTime polymerase chain reaction (RRT-PCR) methods. Bayesian modelling showed similar high sensitivity for the validated H5 HA2 RRT-PCR and a new modified M-gene RRT-PCR that utilizes lyophilized reagents. Both were more sensitive than the validated "wet" M-gene RRT-PCR. Another RRT-PCR, which targeted the H5-gene CS region, was effective for clade 2.3.4 detection, but severely compromised for clade 2.3.2 viruses. Reduced sensitivity of the H5 CS and "wet" M-gene RRT-PCRs correlated with mismatches between the target and the primer and/or probe sequences. However, the H5 HA2 RRT-PCR sensitively detected both clade 2.3.2 and 2.3.4 viruses, and agreed with N1 RRT-PCR results. Feather testing from diseased chicken and duck flocks by AI RRT-PCRs resulted in the most sensitive identification of H5N1 HPAI-infected birds. Evolution of new H5N1 HPAI clades remains a concern for currently affected Asian countries, but also for more distant regions where it is important to be prepared for new incursions of H5N1 HPAI viruses. Genetic evidence for adamantane resistance and sensitivity was also observed in isolates from both clades.

Authors+Show Affiliations

Animal Health and Veterinary Laboratories Agency (AHVLA-Weybridge), Woodham Lane, Addlestone, Surrey, UK. Marek.Slomka@ahvla.gsi.gov.ukNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Comparative Study
Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

22515536

Citation

Slomka, Marek J., et al. "Challenges for Accurate and Prompt Molecular Diagnosis of Clades of Highly Pathogenic Avian Influenza H5N1 Viruses Emerging in Vietnam." Avian Pathology : Journal of the W.V.P.A, vol. 41, no. 2, 2012, pp. 177-93.
Slomka MJ, To TL, Tong HH, et al. Challenges for accurate and prompt molecular diagnosis of clades of highly pathogenic avian influenza H5N1 viruses emerging in Vietnam. Avian Pathol. 2012;41(2):177-93.
Slomka, M. J., To, T. L., Tong, H. H., Coward, V. J., Hanna, A., Shell, W., ... Brown, I. H. (2012). Challenges for accurate and prompt molecular diagnosis of clades of highly pathogenic avian influenza H5N1 viruses emerging in Vietnam. Avian Pathology : Journal of the W.V.P.A, 41(2), pp. 177-93. doi:10.1080/03079457.2012.656578.
Slomka MJ, et al. Challenges for Accurate and Prompt Molecular Diagnosis of Clades of Highly Pathogenic Avian Influenza H5N1 Viruses Emerging in Vietnam. Avian Pathol. 2012;41(2):177-93. PubMed PMID: 22515536.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Challenges for accurate and prompt molecular diagnosis of clades of highly pathogenic avian influenza H5N1 viruses emerging in Vietnam. AU - Slomka,Marek J, AU - To,Thanh L, AU - Tong,Hien H, AU - Coward,Vivien J, AU - Hanna,Amanda, AU - Shell,Wendy, AU - Pavlidis,Theo, AU - Densham,Anstice L E, AU - Kargiolakis,Georgios, AU - Arnold,Mark E, AU - Banks,Jill, AU - Brown,Ian H, PY - 2012/4/21/entrez PY - 2012/4/21/pubmed PY - 2012/8/14/medline SP - 177 EP - 93 JF - Avian pathology : journal of the W.V.P.A JO - Avian Pathol. VL - 41 IS - 2 N2 - Forty-six chickens and 48 ducks were sampled from four Vietnamese poultry premises in 2009 infected with H5N1 highly pathogenic avian influenza (HPAI) clade 2.3.2 and 2.3.4 viruses, which also differed by cleavage site (CS) sequences in their haemagglutinin (HA) genes. All clinical specimens (n=282), namely tracheal and cloacal swabs plus feathers, were tested by five Eurasian reverse-transcriptase AI RealTime polymerase chain reaction (RRT-PCR) methods. Bayesian modelling showed similar high sensitivity for the validated H5 HA2 RRT-PCR and a new modified M-gene RRT-PCR that utilizes lyophilized reagents. Both were more sensitive than the validated "wet" M-gene RRT-PCR. Another RRT-PCR, which targeted the H5-gene CS region, was effective for clade 2.3.4 detection, but severely compromised for clade 2.3.2 viruses. Reduced sensitivity of the H5 CS and "wet" M-gene RRT-PCRs correlated with mismatches between the target and the primer and/or probe sequences. However, the H5 HA2 RRT-PCR sensitively detected both clade 2.3.2 and 2.3.4 viruses, and agreed with N1 RRT-PCR results. Feather testing from diseased chicken and duck flocks by AI RRT-PCRs resulted in the most sensitive identification of H5N1 HPAI-infected birds. Evolution of new H5N1 HPAI clades remains a concern for currently affected Asian countries, but also for more distant regions where it is important to be prepared for new incursions of H5N1 HPAI viruses. Genetic evidence for adamantane resistance and sensitivity was also observed in isolates from both clades. SN - 1465-3338 UR - https://www.unboundmedicine.com/medline/citation/22515536/Challenges_for_accurate_and_prompt_molecular_diagnosis_of_clades_of_highly_pathogenic_avian_influenza_H5N1_viruses_emerging_in_Vietnam_ L2 - http://www.tandfonline.com/doi/full/10.1080/03079457.2012.656578 DB - PRIME DP - Unbound Medicine ER -