Loop-Mediated Amplification for Sensitive and Specific Detection of Viruses

Kalvatchev, Z.; Tsekov, Iliya; Kalvatchev, Nikolay

doi:10.2478/v10133-010-0004-8

Cited by 9 publications

(8 citation statements)

References 15 publications

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“…LAMP is based on auto-cycling strand displacement DNA synthesis by a Bst (Bacillus stearothermophilus) DNA polymerase utilizing four to six primers, which bind laterally to the distinct sites for highly specific amplification in isothermal conditions [26][27][28][29][30][31]. The amplified product from LAMP can be detected using gel electrophoresis, or by adding an intercalating dye (mostly with SYBR™ Green I) to the final end product for visual inspection, or using real-time quantitative measurement by either measuring turbidity using turbidimeter or fluorescence using Genie ® II or Genie ® III (OptiGene, Horsham, WS, UK) [31][32][33][34]. These factors make a LAMP assay suitable to use in laboratory or field conditions.…”

Section: Introductionmentioning

confidence: 99%

Development of Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Cucurbit Leaf Crumple Virus

Waliullah

Ling

Cieniewicz

et al. 2020

IJMS

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A loop-mediated isothermal amplification (LAMP) assay was developed for simple, rapid and efficient detection of Cucurbit leaf crumple virus (CuLCrV), one of the most important begomoviruses that infects cucurbits worldwide. A set of six specific primers targeting a total 240 nt sequence regions in the DNA A of CuLCrV were designed and synthesized for detection of CuLCrV from infected leaf tissues using real-time LAMP amplification with the Genie® III system, which was further confirmed by gel electrophoresis and SYBR™ Green I DNA staining for visual observation. The optimum reaction temperature and time were determined, and no cross-reactivity was seen with other begomoviruses. The LAMP assay could amplify CuLCrV from a mixed virus assay. The sensitivity assay demonstrated that the LAMP reaction was more sensitive than conventional PCR, but less sensitive than qPCR. However, it was simpler and faster than the other assays evaluated. The LAMP assay also amplified CuLCrV-infected symptomatic and asymptomatic samples more efficiently than PCR. Successful LAMP amplification was observed in mixed virus-infected field samples. This simple, rapid, and sensitive method has the capacity to detect CuLCrV in samples collected in the field and is therefore suitable for early detection of the disease to reduce the risk of epidemics.

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Section: Introductionmentioning

confidence: 99%

Development of Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Cucurbit Leaf Crumple Virus

Waliullah

Ling

Cieniewicz

et al. 2020

IJMS

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“…2008). The LAMP technique has been widely used in studies on microbial pathogens, but rarely in animal research (Karanis & Ongerth 2009; Kalvatchev et al. 2010).…”

Section: Discussionmentioning

confidence: 99%

“…The PCR-based method is the main technique employed in genetic studies of animals, but some studies have used LAMP instead because of its high specificity, sensitivity and rapidity under isothermal conditions (Hirayama et al 2006;Nogami et al 2008). The LAMP technique has been widely used in studies on microbial pathogens, but rarely in animal research (Karanis & Ongerth 2009;Kalvatchev et al 2010). In this study, we successfully developed three LAMP reactions for the sex and species identification of Formosa landlocked salmon.…”

Section: Discussionmentioning

confidence: 99%

Species and sex identification of Formosa landlocked salmon using loop‐mediated isothermal amplification

Hsu¹,

Adiputra²,

Ohta³

et al. 2011

Molecular Ecology Resources

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Species and sex identification are among the most important parameters for conservation management. However, it is extremely difficult to perform such identification in Formosa landlocked salmon (Oncorhynchus masou formosanus). Both sexual dimorphism in landlocked dwarf form Formosa landlocked salmon and morphological difference among cherry salmon complex are minimal. We developed a simple, rapid and noninvasive method for identifying sex and species of this critically endangered species using a loop-mediated isothermal amplification (LAMP) technique. The LAMP assay showed the advantage of simple detection (evaluated by visual inspection), rapid reaction time (< 1 h), isothermal condition (less equipment required) and high efficiency (only 0.5-5 pg of DNA was required in the reaction mixture). Therefore, the method is more economical and practical than PCR. The LAMP assay can be easily performed in the field and is a valuable tool for detecting sex ratios in wild populations and identifying species in commercial imports. This is the first application of LAMP in identifying species and sex of salmonids as far as we know and clearly shows the potential application of LAMP in molecular ecology and conservation efforts.

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“…The LAMP reaction can be conducted under isothermal condition ranging from 60 to 65 C [16,17], and specificity is attributable to four primers that recognize six distinct sequences [14][15][16][17][18]. Continuous amplification under isothermal condition produces an extremely large amount of target DNA within less than an hour [7,19], and the method enables simple visual DNA was extracted from positive-MRSA isolates, directly from the samples using boiling method.…”

Section: Introductionmentioning

confidence: 99%

Rapid Detection of Methecillin Resistant Staphylococcus Aureus Using Loop Mediated Isothermal Amplification (LAMP)

Fathelrahman¹,

Hussein²,

Ishag³

et al. 2019

Act Scie Micro

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Background: Staphylococcus aureus, including methicillin-resistant S. aureus (MRSA), is a major bacterial pathogen associated with nosocomial and community-acquired S. aureus infections all over the world. Aim: The aim of this study was to establish the loop mediated isothermal amplification technique as a rapid detection method for methicillin resistant S. aureus. Methods: The study was carried out in Omdurman Military hospital, Alshorta hospital, and Alneelain university dental clinic. A total of 60 samples were collected and cultured, All confirmed S. aureus isolates were tested against oxacillin antibiotic by the disk diffusion method to detect MRSA isolates LAMP and PCR assays were then used to detect mec A gene directly from the samples and from culture isolates. Results: Among 60 (wound, dental plaque and urine) seven samples (11.6%) MRSA were detected by culture, 5% were found positive for mec A using LAMP and 3.3% using PCR. LAMP detected mec A in 100% of the seven MRSA isolates, PCR detected mec A in 14.3%. The LAMP results were confirmed using melting analysis. Conclusion: The LAMP described herein is a reliable and rapid method for detection of the mec A gene. Generally, these findings are useful for future studies since there is little available information about MRSA infection in Sudan. LAMP can be used in a hospital setting for rapid diagnosis of MSRA, this should help better management and treatment of the affected patients in addition to rapid initiation of infection control procedures.

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Loop-Mediated Amplification for Sensitive and Specific Detection of Viruses

Cited by 9 publications

References 15 publications

Development of Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Cucurbit Leaf Crumple Virus

Development of Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Cucurbit Leaf Crumple Virus

Species and sex identification of Formosa landlocked salmon using loop‐mediated isothermal amplification

Rapid Detection of Methecillin Resistant Staphylococcus Aureus Using Loop Mediated Isothermal Amplification (LAMP)

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