Differential detection of Wheat yellow mosaic virus, Japanese soil-borne wheat mosaic virus and Chinese wheat mosaic virus by reverse transcription loop-mediated isothermal amplification reaction

Fukuta, Shiro; Tamura, Masahito; Hidekazu, Maejima; Takahashi, Reiko; Kuwayama, Sachiko; Tsuji, Takako; Yoshida, Tomokazu; Itoh, Koji; Hashizume, Hajime; Nakajima, Yasunori; Uehara, Yasushi; Shirako, Yukio

doi:10.1016/j.jviromet.2013.03.005

Cited by 46 publications

(16 citation statements)

References 31 publications

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“…This assay was carried out in triplicate for each template DNA. Previous studies suggest that the LAMP assay has the same or higher sensitivity than the PCR assay (Fukuta et al ., ). In this study, the sensitivity of the LAMP assay was equivalent to that of conventional PCR (Ishiguro et al ., ).…”

Section: Resultsmentioning

confidence: 97%

Development and application of a loop-mediated isothermal amplification assay for rapid detection ofPythium helicoides

Takahashi¹,

Fukuta²,

Kuroyanagi³

et al. 2014

FEMS Microbiol Lett

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Root rot of poinsettia, caused by Pythium helicoides at high temperatures in hydroponic cultures, has become a serious problem in many parts of the world. We have developed a species-specific, loop-mediated isothermal amplification (LAMP) assay for the rapid diagnosis of this pathogen. The primers were designed using the ribosomal DNA internal transcribed spacer sequence. Primer specificity was established using 40 Pythium species including P. helicoides, 11 Phytophthora species, and eight other soil-borne pathogens. A sensitivity test was carried out using genomic DNA extracted from P. helicoides, and the detection limit was c. 100 fg which is comparable to that of the polymerase chain reaction (PCR). In addition, we tested the ease of pathogen detection in poinsettia roots. The LAMP results were consistent with those from the conventional plating method and showed more sensitivity than the PCR results. Consequently, the LAMP method developed in this study is effective for the rapid and easy detection of P. helicoides.

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Section: Resultsmentioning

confidence: 97%

Development and application of a loop-mediated isothermal amplification assay for rapid detection ofPythium helicoides

Takahashi¹,

Fukuta²,

Kuroyanagi³

et al. 2014

FEMS Microbiol Lett

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“…LAMP products amplified by different LAMP primer sets differ in peak temperature (Fukuta et al . ,b). Therefore, monitoring denaturing peak temperatures helps distinguishing LAMP products without electrophoresis.…”

Section: Resultsmentioning

confidence: 99%

“…; Fukuta et al . ,b). LAMP requires four different primers designed specifically to recognize six distinct regions on the target gene.…”

Section: Introductionmentioning

confidence: 99%

Development of loop-mediated isothermal amplification assay for the detection of Pythium myriotylum

Fukuta¹,

Takahashi²,

Kuroyanagi³

et al. 2014

Lett Appl Microbiol

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Significance and Impact of the Study: This study shows the first LAMP assay for the detection of Pythium myriotylum. The primer set designed from ITS region of P. myriotylum can detect the pathogen in field sample with a fast and convenient method. Analysis of the annealing curve of the LAMP reaction products increases the reliability of the LAMP diagnosis. This study shows that the diagnostic method using the LAMP assay is useful for monitoring P. myriotylum in the field. AbstractThis study reports the development of a loop-mediated isothermal amplification (LAMP) reaction for the detection of Pythium myriotylum. The primer set targeting the ITS sequence of P. myriotylum worked most efficiently at 60°C and allowed the detection of P. myriotylum DNA within 30 min by fluorescence monitoring using a real-time PCR instrument. The peak denaturing temperature of amplified DNA was about 87Á0°C. In specificity tests using eight Pythium myriotylum strains, 59 strains from 39 species of Pythium, 11 Phytophthora strains and eight other soil-borne pathogens, LAMP gave no cross-reactions. The detection limit was 100 fg of genomic DNA, which was as sensitive as PCR. LAMP could detect P. myriotylum in hydroponic solution samples, and the results coincided with those of the conventional plating method in almost all cases. The LAMP method established in this study is a simple and sensitive tool for the detection of P. myriotylum.

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“…There are several methods for analysing the mLAMP assay further, such as measuring the fluorescent signal of the LAMP reactions using a real‐time fluorometer (Dong, Cho, Hahn, & Cho, ; Fukuta et al., ; Tanner, Zhang, & Evans, ) and analysing the amplified bands for polymorphisms using restriction enzymes and electrophoresis (Iseki et al., ; Shao, Zhu, Jin, & Chen, ). Although these methods are useful, they have a number of limitations.…”

Section: Discussionmentioning

confidence: 99%

Multiplex loop‐mediated isothermal amplification assay for the identification of three major whitefly species in the greenhouse

Suzuki¹,

Tanaka²,

Suzuki³

et al. 2018

J Applied Entomology

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A multiplex loop‐mediated isothermal amplification (mLAMP) assay was developed for the identification of three species of whitefly, Trialeurodes vaporariorum, Bemisia tabaci Middle East‐Asia Minor 1 (MEAM1) and Mediterranean (MED), major pests in the greenhouse. Each of the specific LAMP primer sets was designed based on the mitochondrial cytochrome oxidase I (mtCOI) gene sequence. The mLAMP reactions using primer mixtures labelled with fluorescent dye were performed at 63°C for 60 min and centrifuged with polyethyleneimine. Thus, T. vaporariorum, MEAM1 and MED were clearly identified by the colour precipitates under UV light. The mLAMP procedure described in this study is cost‐effective and can be performed in the field not only in the laboratory, because this method is a single analysis and does not need a special gene amplification device.

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Differential detection of Wheat yellow mosaic virus, Japanese soil-borne wheat mosaic virus and Chinese wheat mosaic virus by reverse transcription loop-mediated isothermal amplification reaction

Cited by 46 publications

References 31 publications

Development and application of a loop-mediated isothermal amplification assay for rapid detection ofPythium helicoides

Development and application of a loop-mediated isothermal amplification assay for rapid detection ofPythium helicoides

Development of loop-mediated isothermal amplification assay for the detection of Pythium myriotylum

Multiplex loop‐mediated isothermal amplification assay for the identification of three major whitefly species in the greenhouse

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