Development of H5-RT-LAMP (loop-mediated isothermal amplification) system for rapid diagnosis of H5 avian influenza virus infection

Imai, Masaki; Ninomiya, Ai; Minekawa, Harumi; Notomi, Tsugunori; Ishizaki, Takayoshi; Tashiro, Masato; Odagiri, Takato

doi:10.1016/j.vaccine.2006.05.046

Cited by 159 publications

(120 citation statements)

References 7 publications

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“…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.…”

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confidence: 99%

“…The extremely high variability of the AIV hemagglutinin (HA) gene as a target for HA subtype-specific diagnostics poses a particularly difficult case not only for the design of primers for conventional PCR, but even more for appropriate LAMP primer sets. 5,12,13 The need for new rapid AIV assays that can be carried out with a minimum of specific equipment prompted the present evaluation of the performance characteristics of a commercial real-time RT-LAMP amplification kit a and the corresponding primer kits a designed to detect RNA of the notifiable H5-and H7-AIV subtypes. 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.…”

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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.…”

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confidence: 99%

“…Concerning sensitivities of AIV subtype-specific LAMP approaches, only a few reports on H5 and 1 report on H9 have been published so far, but these focus more on the suitability for detection of a selection of very recent strains rather than a wide variety of distinct isolates. 2,5,8 Surprisingly, no data is available for the diagnosis of H7 subtype AIV RNA with the use of LAMP-based assays, although H7 AIV infections of poultry are notifiable to the O.I.E (World Organization for Animal Health). 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.…”

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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.…”

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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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confidence: 99%

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confidence: 99%

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confidence: 99%

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confidence: 99%

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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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Loop‐mediated isothermal amplification‐based diagnostic assay for monkeypox virus infections

Iizuka

Saijo

Shiota

et al. 2009

Journal of Medical Virology

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Monkeypox virus (MPXV) causes a smallpox-like disease in non-human primates and humans. This infection is endemic to central and western Africa. MPXV is divided into two genetically different groups, Congo Basin and West African MPXV, with the former being the more virulent. A real-time quantitative MPXV genome amplification system was developed for the diagnosis of MPXV infections using loop-mediated isothermal amplification (LAMP) technology. Primers used for genome amplification of Congo Basin (C-LAMP), West African (W-LAMP), and both Congo Basin and West African (COM-LAMP) MPXV by LAMP were designed according to the nucleotide sequences of the Congo Basin-specific D14L gene, the West African-specific partial ATI gene, and the partial ATI gene that is shared by both groups, respectively. The sensitivity and specificity of the LAMP were evaluated with nested PCR using peripheral blood and throat swab specimens collected from Congo Basin MPXV or West African MPXV-infected monkeys. The sensitivity and specificity of COM-LAMP, C-LAMP, and W-LAMP were 80% (45/56) and 100% (64/64); 79% (19/24) and 100% (24/24); and 72% (23/32) and 100% (40/40), respectively. The viremia level determined by LAMP assays increased with increases in the severity of the monkeypox-associated symptoms. The newly developed LAMP assay was confirmed to be a rapid, quantifiable, and highly sensitive and specific system effective in the diagnosis of MPXV infections. The LAMP assays made it possible to discriminate between Congo Basin and West African MPXV. The LAMP developed in this study is useful not only for diagnosis of but also for the assessment of MPXV infections.

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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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Development of H5-RT-LAMP (loop-mediated isothermal amplification) system for rapid diagnosis of H5 avian influenza virus infection

Cited by 159 publications

References 7 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

Loop‐mediated isothermal amplification‐based diagnostic assay for monkeypox virus infections

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

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