A NASBA Method to Detect High- and Low-Pathogenicity H5 Avian Influenza Viruses

Collins, Richard A.; Ko, Lung-Sang; So, Ka-Lun; Ellis, T.M.; Lau, Lok Ting; Yu, Albert Cheung-Hoi

doi:10.1637/0005-2086-47.s3.1069

Cited by 17 publications

(14 citation statements)

References 6 publications

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“…These detect H5 HA specific sequence of virus RNA with H5 HA gene specific primer pairs (7,12,21), or detect antibodies to H5 HA of influenza virus (9,22). However, it takes several hours to obtain results and specific reagents and instruments are needed.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Development of an Immunochromatographic Kit for Rapid Diagnosis of H5 Avian Influenza Virus Infection

Tsuda

Sakoda

Sakabe

et al. 2007

Microbiology and Immunology

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Highly pathogenic avian influenza (HPAI) caused by the H5N1 subtype has given rise to serious damage in poultry industries in Asia. The virus has expanded its geographical range to Europe and Africa, posing a great risk to human health as well. For the control of avian influenza, a rapid diagnosis by detecting the causative virus and identifying its subtype is essential. In the present study, a rapid diagnosis kit combining immunochromatography with enzyme immunoassay which detects the H5 HA antigen of influenza A virus was developed using newly established anti-H5 HA monoclonal antibodies. The present kit specifically detected all of the H5 influenza viruses tested, and did not react with the other HA subtypes. H5 HA antigens were detected from swabs and tissue homogenates of chickens infected with HPAI virus strain A/chicken/Yamaguchi/7/04 (H5N1) from 2 days post inoculation. The kit showed enough sensitivity and specificity for the rapid diagnosis of HPAI.

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Section: Discussionmentioning

confidence: 99%

“…Rapid diagnosis kits are available to detect influenza virus specific nucleoprotein (NP) of which antigenicity is common among the A or B type influenza viruses, but do not identify the HA subtype of influenza A virus (1,4 (7,18).…”

mentioning

confidence: 99%

Development of an Immunochromatographic Kit for Rapid Diagnosis of H5 Avian Influenza Virus Infection

Tsuda

Sakoda

Sakabe

et al. 2007

Microbiology and Immunology

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“…In 2004, Lau et al used this method to detect a portion of the HA gene of avian influenza A virus subtypes H5 and H7 irrespective of lineage and also used the matrix gene to detect all the then known subtypes (H1–H15) of AIV [58]. NASBA methods have also been used for surveillance of low pathogenicity and HPAI H5 AIVs [59]. In a recent study, a real-time NASBA was used for molecular detection of influenza A H1N1 and H3N2, influenza B, respiratory syncytial virus and human metapneumovirus using human respiratory samples collected on dry cotton swabs [60].…”

Section: Nucleic Acid Sequence-based Methodsmentioning

confidence: 99%

Methods for molecular surveillance of influenza

Wang

Taubenberger

2010

Expert Review of Anti-infective Therapy

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Molecular-based techniques for detecting influenza viruses have become an integral component of human and animal surveillance programs in the last two decades. The recent pandemic of the swineorigin influenza A virus (H1N1) and the continuing circulation of highly pathogenic avian influenza A virus (H5N1) further stress the need for rapid and accurate identification and subtyping of influenza viruses for surveillance, outbreak management, diagnosis and treatment. There has been remarkable progress on the detection and molecular characterization of influenza virus infections in clinical, mammalian, domestic poultry and wild bird samples in recent years. The application of these techniques, including reverse transcriptase-PCR, real-time PCR, microarrays and other nucleic acid sequencing-based amplifications, have greatly enhanced the capability for surveillance and characterization of influenza viruses.

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“…Efficient transcription of full-length products is required for many classes of assays, such as microarray analysis and NASBA techniques. NASBA assays generally limit the size of the amplified product to less than 600 bases, and many are significantly shorter [40,41]. For the influenza genome, both ends are highly conserved as required by the native viral polymerase for a priming site, but the rest of the sequence is highly variable.…”

Section: Impact For Diagnostic Rna Synthesismentioning

confidence: 99%

Early In Vitro Transcription Termination in Human H5 Influenza Viral RNA Synthesis

Kerby

Sarma

Patel

et al. 2011

Appl Biochem Biotechnol

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Rapid diagnostic identification of the human H5 influenza virus is a strategic cornerstone for outbreak prevention. We recently reported a method for direct detection of viral RNA from a highly pathogenic human H5 influenza strain (A/Hanoi/30408/2005(H5N1)), which necessarily was transcribed in vitro from non-viral sources. This article provides an in-depth analysis of the reaction conditions for in vitro transcription (IVT) of full-length influenza H5 RNA, which is needed for diagnostic RNA production, for the T7 and SP6 phage promoter systems. Gel analysis of RNA transcribed from plasmids containing the H5 sequence between a 5' SP6 promoter and 3' restriction site (BsmBI) showed that three sequence-verified bands at 1,776, 784, and 591 bases were consistently produced, whereas only one 1,776-base band was expected. These fragments were not observed in H1 or H3 influenza RNA transcribed under similar conditions. A reverse complement of the sequence produced only a single band at 1,776 bases, which suggested either self-cleavage or early termination. Aliquots of the IVT reaction were quenched with EDTA to track the generation of the bands over time, which maintained a constant concentration ratio. The H5 sequence was cloned with T7 and SP6 RNA polymerase promoters to allow transcription in either direction with either polymerase. The T7 transcription product from purified, restricted plasmids in the vRNA direction only produced the 1,776-base full-length sequence and the 784-base fragment, instead of the three bands generated by the SP6 system, suggesting an early termination mechanism. Additionally, the T7 system produced a higher fraction of full-length vRNA transcripts than the SP6 system did under similar reaction conditions. By sequencing we identified a type II RNA hairpin loop terminator, which forms in a transcription direction-dependent fashion. Variation of the magnesium concentration produced the greatest impact on termination profiles, where some reaction mixtures were unable to produce full-length transcripts. Optimized conditions are presented for the T7 and SP6 phage polymerase systems to minimize these early termination events during in vitro transcription of H5 influenza vRNA.

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A NASBA Method to Detect High- and Low-Pathogenicity H5 Avian Influenza Viruses

Cited by 17 publications

References 6 publications

Development of an Immunochromatographic Kit for Rapid Diagnosis of H5 Avian Influenza Virus Infection

Development of an Immunochromatographic Kit for Rapid Diagnosis of H5 Avian Influenza Virus Infection

Methods for molecular surveillance of influenza

Early In Vitro Transcription Termination in Human H5 Influenza Viral RNA Synthesis

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