Rapid diagnosis of H5N1 avian influenza virus infection by newly developed influenza H5 hemagglutinin gene-specific loop-mediated isothermal amplification method

Imai, Masaki; Ninomiya, Ai; Minekawa, Harumi; Notomi, Tsugunori; Ishizaki, Takayoshi; Tú, Phan Văn; Tien, Nguyen Thi Kim; Tashiro, Masato; Odagiri, Takato

doi:10.1016/j.jviromet.2006.12.004

Cited by 130 publications

(109 citation statements)

References 22 publications

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“…It had been reported previously that, for some of the older H5 isolates like A/tern/ South Africa/61, LAMP-based RNA detection is about 1,000 times less sensitive, even compared with conventional RT-PCR. 5,6 This observation is supported by the finding that this isolate also owned the lowest detection limits in the evaluated H5 real-time RT-LAMP. Real-time RT-PCR proved to be even 100,000 times more sensitive in this case.…”

supporting

confidence: 61%

“…Problems of designing an optimally matching universal set of broadly reactive H5-or H7-specific LAMP primers are inherent in the high variability of the Influenza A virus HA sequences and the special constraints of LAMP primer design. 5,6 The observed lower average sensitivity of the H5-and H7-specific real-time RT-LAMP assays in our study might be due to primers that do not equally well match with the phylogenetically very divergent isolates used in this broad evaluation. LAMP primers tailored to detect with high sensitivity a limited set of, e.g., H5 isolates currently circulating in a circumscribed region are expected to yield better sensitivities.…”

Section: Virus Isolatementioning

confidence: 78%

“…5,6 Specific H5N1 virus isolates of Asian origin belonging to different phylogenetic clades, as well as some Eurasian LPAI H5 viruses, were particularly well detected (Table 1, indicated by boldface). However, a number of H5 isolates, including several Southeast Asian HPAIV strains, as well as LPAIV isolates of recent Eurasian origin, were detected with diminished sensitivity.…”

mentioning

confidence: 99%

“…Until then, no data were available concerning performance characteristics of the H7-specific real-time RT-LAMP, whereas H5-specific kits had been evaluated with some selected virus isolates of the HPAIV H5N1 and were found to have a matching, if not superior, performance when compared with conventional RT-PCR. 5,6 However, real-time RT-PCR is considerably more sensitive than conventional RT-PCR and now represents the gold standard for AIV RNA detection, 12 and no data on comparisons between RT-LAMP and subtypespecific real-time RT-PCR are available.…”

mentioning

confidence: 99%

“…All AIV-targeted RT-LAMP assays developed so far have been compared with conventional RT-PCR, and sensitivity was calculated on the basis of infectivity, not taking into account the amount of RNA copies. 2,5,6,8 In this study, sensitivity of real-time RT-LAMP assays was compared with highly sensitive realtime RT-PCR assays, which now represent the gold standard for AIV RNA detection. 12 A conventional RT-LAMP assay developed for detection of the nucleoprotein gene of human Influenza B virus and the HA fragments of the Influenza A virus subtypes H1 and H3 was reported to be less sensitive than virus isolation but more sensitive than a commercial immunochromatography test.…”

mentioning

confidence: 99%

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Evaluation of Two Commercial Loop-Mediated Isothermal Amplification Assays for Detection of Avian Influenza H5 and H7 Hemagglutinin Genes

Postel

Letzel

Frischmann³

et al. 2010

J VET Diagn Invest

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Abstract. Real-time reverse transcription loop-mediated isothermal amplification (real-time RT-LAMP) holds substantial potential as a highly sensitive, specific, and easy-to-perform molecular technique for pathogen detection in clinical samples. In the current study, the analytical and diagnostic performance of 2 commercial realtime RT-LAMP kits, Avian Flu H5 and Avian Flu H7, in detecting Avian influenza virus (AIV) infections were evaluated and compared with validated real-time reverse transcription polymerase chain reaction (RT-PCR) assays using RNA from reference virus isolates of subtypes H5 (n 5 24) and H7 (n 5 25) and of phylogenetically related subtypes (n 5 20). When real-time RT-LAMP was carried out according to the recommendations of the manufacturer, 3 out of 24 H5 isolates and 8 out of 25 H7 reference strains were not detected. Prolonging the amplification phase resulted in detection of all H5 isolates but also in false positive detection of 2 non-H5 isolates. Real-time RT-LAMP specific to H7 failed to detect 2 H7 isolates after prolonged amplification. According to the examination of RNA log dilutions, the sensitivity of the real-time RT-LAMP assays, for a number of historic but also recent strains, was considerably lower compared with subtype-specific real-time RT-PCR assays. Application of the real-time RT-LAMP assays for analysis of diagnostic samples from wild birds confirmed their lower sensitivity. Commercial real-time RT-LAMP as tested in this study with a broad range of AIV H5 and H7 strains of phylogenetically diverse yet recent origin, holds some promise for routine veterinary diagnostic purposes, although real-time RT-LAMP was markedly more vulnerable to a reduction of detection limits because of strainspecific sequence variation than subtype-specific real-time RT-PCR.

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supporting

confidence: 61%

Section: Virus Isolatementioning

confidence: 78%

mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Evaluation of Two Commercial Loop-Mediated Isothermal Amplification Assays for Detection of Avian Influenza H5 and H7 Hemagglutinin Genes

Postel

Letzel

Frischmann³

et al. 2010

J VET Diagn Invest

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Point‐of‐Care Diagnostic Platforms for Loop‐Mediated Isothermal Amplification

et al. 2022

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The loop‐mediated isothermal amplification (LAMP) method is one of the Nucleic acid amplification tests (NAATs) that allows for the amplification of target regions without using a thermal cycle. With its unique primer design, LAMP ensures the rapid replication of the targeted DNA region with high specificity and high efficiency. LAMP technology is used for diagnostic purposes in pathogen detection due to its ease of use, low cost, and simplicity without requiring complex equipment. A wide range of LAMP diagnostic platforms have been developed for applications in bacteria, virus, and parasitic pathogen detection. Herein, the methodology of LAMP technology and its applications in pathogen detection and SNP genotyping and mutation detection are discussed. Point‐of‐care (PoC) LAMP platforms designed with the principles of microfluidic chip technology, including LAMP‐on‐a‐chip, paper‐based LAMP, and smartphone‐based LAMP applications have been elaborated. LAMP technology represents a fast, robust, and reliable diagnostic platform for point‐of‐care testing.

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Evaluation of reverse transcription loop‐mediated isothermal amplification assays for rapid diagnosis of pandemic influenza A/H1N1 2009 virus

Nakauchi

Yoshikawa

Nakai

et al. 2010

Journal of Medical Virology

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Two genetic diagnosis systems using reverse transcription-loop-mediated isothermal amplification (RT-LAMP) technology were evaluated: one for detecting the HA gene of the pandemic influenza A/H1N1 2009 virus (H1pdm RT-LAMP) and the other for detecting the matrix gene of the influenza A virus (TypeA RT-LAMP). The competence of these two RT-LAMP assay kits for the diagnosis of the pandemic influenza A/H1N1 2009 virus was compared using real-time RT-PCR assays developed recently on viruses isolated and clinical specimens collected from patients with suspected infection. TypeA RT-LAMP and H1pdm RT-LAMP showed almost the same sensitivity as real-time RT-PCR for viruses isolated. The sensitivity and specificity of TypeA RT-LAMP and H1pdm RT-LAMP were 96.3% and 88.9%, respectively, for clinical specimens. Considering that the ability of the two RT-LAMP assay kits for detection of the pandemic influenza A/H1N1 2009 virus was comparable to that of the real-time RT-PCR assays, and that the assays were completed within 1 hr and did not require any expensive equipment, these two RT-LAMP assays are promising rapid diagnostic tests for the pandemic influenza A/H1N1 2009 virus at the hospital bedside.

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Rapid diagnosis of H5N1 avian influenza virus infection by newly developed influenza H5 hemagglutinin gene-specific loop-mediated isothermal amplification method

Cited by 130 publications

References 22 publications

Evaluation of Two Commercial Loop-Mediated Isothermal Amplification Assays for Detection of Avian Influenza H5 and H7 Hemagglutinin Genes

Evaluation of Two Commercial Loop-Mediated Isothermal Amplification Assays for Detection of Avian Influenza H5 and H7 Hemagglutinin Genes

Point‐of‐Care Diagnostic Platforms for Loop‐Mediated Isothermal Amplification

Evaluation of reverse transcription loop‐mediated isothermal amplification assays for rapid diagnosis of pandemic influenza A/H1N1 2009 virus

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