Novel reverse transcription loop-mediated isothermal amplification for rapid detection of foot-and-mouth disease virus

Dukes, Juliet; King, Donald P.; Alexandersen, Søren

doi:10.1007/s00705-005-0708-5

Cited by 212 publications

(187 citation statements)

References 17 publications

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“…During the present study, the application of a constant temperature of 65 °C for 60 min is similar to Dukes et al (14). Moreover, RT-LAMP was found suitable for FMDV serotyping at 63 °C for 60 min; this finding was also reported by Madhanmohan et al (9).…”

Section: Discussionsupporting

confidence: 90%

“…Furthermore, the specificity of RT-LAMP was 100% with no cross-reactivity and this was also observed by Madhanmohan et al (9). In contrast to this, Dukes et al (14) evaluated RT-LAMP assay with TaqMan rRT-PCR and reported 81/98 (82.65%) and 94/98 (95.91%) positive by RT-LAMP and rRT-PCR, respectively. However, Dukes et al (14) reported that this may be due to high genetic variation among the FMDV genome.…”

Section: Discussionmentioning

confidence: 55%

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One-step real-time loop-mediated isothermal amplification (RT-LAMP): evaluation and its application for the detection of foot-and-mouth-disease virus and its serotypes

Maryam¹,

Rasheed²,

Latif³

et al. 2017

Turk J Vet Anim Sci

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Availability of a rapid and sensitive diagnostic technique is key in successful prevention and control of infectious diseases like foot-and-mouth disease (FMD). Existing conventional diagnostic tests for FMD are laborious and time-consuming with low sensitivity and specificity. Molecular-based techniques are costly and difficult, involving refined apparatus like a thermal cycler. In the present study, the technique of real-time loop-mediated isothermal amplification (RT-LAMP) was standardized for diagnosing FMDV and its serotypes, evaluated using field samples, and compared with the existing real-time PCR in Pakistan. RT-LAMP amplified the target 3D gene using specific primers at 65 °C for 60 min and the VP1 gene using serotype specific primers at 63 °C for 60 min. A total of 38 samples out of 50 were positive by RT-LAMP and identified serotypes were A (n = 15), O (n = 15), and Asia-1 (n = 8). The efficiency of RT-LAMP in this study was highest for serotype Asia-1 (80.9%) followed by serotype A (73.4%) and O (62.35).

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Section: Discussionsupporting

confidence: 90%

Section: Discussionmentioning

confidence: 55%

One-step real-time loop-mediated isothermal amplification (RT-LAMP): evaluation and its application for the detection of foot-and-mouth-disease virus and its serotypes

Maryam¹,

Rasheed²,

Latif³

et al. 2017

Turk J Vet Anim Sci

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“…Gradual depletion of magnesium ion during amplification reaction is the reason behind change of pre-amplification violet colour of reaction mixture to post-amplification sky blue colour in a positive LAMP reaction [6]. DNA intercalating dyes like SYBR green I and picogreen can be used for detecting amplified product [19,20]. But it requires a UV light source for visualisation.…”

Section: Resultsmentioning

confidence: 99%

Visual Detection of Brucella spp. in Spiked Bovine Semen Using Loop-Mediated Isothermal Amplification (LAMP) Assay

et al. 2016

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Several pathogens including Brucella spp. are shed in semen of infected bulls and can be transmitted to cows through contaminated semen during artificial insemination. The present study reports omp2a and bcsp31 gene based loop-mediated isothermal amplification (LAMP) assays for detection of Brucella genomic DNA in semen from infected bulls. The positive results could be interpreted visually by change in colour of reaction mixture containing hydroxyl naphthol blue (HNB) dye from violet to sky blue. LAMP assays based on omp2a and bcsp31 could detect as little as 10 and 100 fg of B. abortus S19 genomic DNA, respectively. Sensitivity of omp2a and bcsp31 LAMP assays for direct detection of organisms in bovine semen was 2.28 9 10 1 CFU and 2.28 9 10 2 CFU of B. abortus S19 in spiked bovine semen, respectively. The omp2a LAMP assay was found equally sensitive to TaqMan probe based real-time PCR and 100 times more sensitive than conventional PCR in identifying Brucella in spiked semen. The diagnostic applicability of the omp2a LAMP assay was evaluated with seventy-nine bovine semen samples and results were re-evaluated through TaqMan probe based real-time PCR and conventional PCR. Taken together, the omp2a LAMP assay is easy to perform, rapid and sensitive in diagnosis of Brucella spp. in bovine semen.

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“…The results of LAMP can be observed directly with the naked eye after adding an intercalating dye and visualised by gel-electrophoresis or real-time fluorogenic analysis with a thermal cycler. LAMP assays have been used in detecting a variety of pathogens, including animal viruses (7,9). A set of four primers targeting conserved segment 1 of BTV RNA was used in an RT-LAMP reaction developed in our laboratory for the detection of BTV genome (30).…”

mentioning

confidence: 99%

Molecular technologies for detection and typing of bluetongue virus

Niedbalski

2017

Medycyna Weterynaryjna

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Rapid and accurate diagnosis plays an important role in the implementation of effective measures to control the spread of disease. Historically, the laboratory diagnosis and typing of BTV were carried out by various serological and virological methods, including virus neutralization (VN) assay, ELISA, as well as virus isolation (VI) in cell cultures or in embryonated chicken eggs. At present, various molecular techniques to detect BTV genome are increasingly used as primary diagnostic tools for the serotyping and epidemiological investigations of BTV. Initially, the viral RNA was detected by simple nucleic acid hybridization technologies. Then, conventional RT-PCR assays were developed and evaluated for the detection of BTV serotypes based on nucleotide sequences of different genome segments. Although RT-PCR, with its increased sensitivity, has advantages over hybridization, it is almost impossible to quantify accurately by regular and multiplex PCR procedures, and regular PCR may produce false positive results. Over the recent years, a number of real-time RT-PCR (rRT-PCR) methods have been described. The rRT-PCR offers certain advantages over conventional RT-PCR assay, as it is more rapid, sensitive, and can provide quantitative as well as qualitative genetic information. It does not use agarose gel electrophoresis, decreases the risk of contamination because it is run within an enclosed tube, and is suitable for large-scale testing and automation. The target amplicon is usually smaller, reducing the potential for problems caused by target degradation. Loop-mediated isothermal amplification (LAMP), a novel rapid, accurate and cost effective gene amplification method, is an autocycling and strand displacement DNA synthesis method. LAMP assays have been applied as a method of detecting a variety of animal pathogens, including BTV. RT- LAMP assay can be a valuable tool complementing the routine laboratory diagnosis of BTV.

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Novel reverse transcription loop-mediated isothermal amplification for rapid detection of foot-and-mouth disease virus

Cited by 212 publications

References 17 publications

One-step real-time loop-mediated isothermal amplification (RT-LAMP): evaluation and its application for the detection of foot-and-mouth-disease virus and its serotypes

One-step real-time loop-mediated isothermal amplification (RT-LAMP): evaluation and its application for the detection of foot-and-mouth-disease virus and its serotypes

Visual Detection of Brucella spp. in Spiked Bovine Semen Using Loop-Mediated Isothermal Amplification (LAMP) Assay

Molecular technologies for detection and typing of bluetongue virus

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