Loop-mediated isothermal amplification (LAMP): principle, features, and future prospects

Notomi, Tsugunori; Mori, Yasuyoshi; Tomita, Norihiro; Kanda, Hidetoshi

doi:10.1007/s12275-015-4656-9

Cited by 698 publications

(525 citation statements)

References 14 publications

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“…The assay uses primers designed to target the capsid gene and generate real-time amplification that can be detected in less than 30 min. These characteristics make LAMP assays ideal to use in portable devices, which can be configured to diagnose diseases at point of care facilities [14,15]. Therefore, the detection of ZIKV RNA by LAMP provides the basis of a test that can readily facilitate the diagnosis of new cases.…”

Section: Introductionmentioning

confidence: 99%

Rapid diagnosis of Zika virus through saliva and urine by Loop-mediated isothermal amplification (LAMP)

Castro

Sabalza

Barber

et al. 2018

Journal of Oral Microbiology

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Background: Zika virus (ZIKV) is a single-stranded RNA virus and member of the Flaviviridae family. Recent studies have reported that saliva can be an important alternative to detect ZIKV. Saliva requires less processing than blood greatly simplifying the assay. Loop-mediated Isothermal Amplification (LAMP) is a rapid assay that detects nucleic acids, including ZIKV RNA. Aim: The aim of this study was to evaluate the efficacy of saliva and urine to diagnose ZIKV infection in subjects during the acute phase, through ZIKV RNA detection by LAMP. Method: A total of 131 samples (68 saliva and 63 urine samples) from 69 subjects in the acute phase of ZIKV infection, and confirmed positive for ZIKV by blood analysis through real time-PCR, were collected and analyzed by Reverse Transcriptase Loop-mediated Isothermal Amplification (RT-LAMP). Results: From the 68 saliva samples, 45 (66.2%) were positive for ZIKV with an average time to positivity (Tp) of 13.5 min, and from the 63 urine samples, 25 (39.7%) were positive with the average Tp of 15.8 min. Saliva detected more samples (p = 0.0042) and had faster Tp (p = 0.0176) as compared with urine. Conclusion: Saliva proved to be a feasible alternative to diagnose ZIKV infection during the acute phase by LAMP.

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Section: Introductionmentioning

confidence: 99%

Rapid diagnosis of Zika virus through saliva and urine by Loop-mediated isothermal amplification (LAMP)

Castro

Sabalza

Barber

et al. 2018

Journal of Oral Microbiology

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“…LAMP employs only a single type of DNA polymerase with high strand displacement activity that amplifies DNA at a constant temperature. [33] A set of four specially designed primers (two inner (FIB/BIP) and two outer primers (F3/B3)) targeting six distinct gene regions are used for amplification. The methodology of amplification relies on repetition of two types of elongation reactions occurring at the loop regions: self-elongation of templates from the stem loop structure formed at the 3 0 -terminal and the binding and elongation of new primers from the loop region.…”

Section: Loop-mediated Isothermal Amplification (Lamp)mentioning

confidence: 99%

“…The elongation reactions are then sequentially repeated using the stem-loop region as a template with both inner primers and DNA polymerase-mediated strand-displacement synthesis. [33] The final products of the amplification are a mix of stem-loop DNA structures of different lengths and cauliflower-like structures formed by annealing between alternatively inverted repeats of the target in the same strand. This method can amplify a few copies of DNA to 10 9 in less than an hour at constant temperature and with great specificity due to the fundamental characteristics of the inner primers.…”

Section: Loop-mediated Isothermal Amplification (Lamp)mentioning

confidence: 99%

Loop‐mediated isothermal amplification (LAMP) as an alternative to PCR: A rapid on‐site detection of gene doping

Salamin

Kuuranne

Saugy

et al. 2017

Drug Testing and Analysis

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Innovation in medical research has been diverted at multiple occasions to enhance human performance. The predicted great progress in gene therapy has raised some concerns regarding its misuse in the world of sports (gene doping) for several years now. Even though there is no evidence that gene doping has ever been used in sports, the continuous improvement of gene therapy techniques increases the likelihood of abuse. Therefore, since 2004, efforts have been invested by the anti-doping community and WADA for the development of detection methods. Several nested PCR and qPCR-based strategies exploiting the absence of introns in the transgenic DNA have been proposed for the long-term detection of transgene in blood. Despite their great sensitivity, those protocols are hampered by limitations of the techniques that can be cumbersome and costly. The purpose of this perspective is to describe a new approach based on loop-mediated isothermal amplification (LAMP) for the detection of gene doping. This protocol enables a rapid and simple method to amplify nucleic acids with a high sensitivity and specificity and with a simple visual detection of the results. LAMP is already being used in clinical application for the detection of viruses or mutations. Therefore, this technique has the potential to be further developed for the detection of foreign genetic material in elite athletes.

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“…In 2000, a novel nucleic acid amplification method known as loop-mediated isothermal amplification (LAMP) was developed (Notomi et al, 2000), and was considered a breakthrough technique for the diagnosis of TB and other infectious diseases (Mori & Notomi, 2009;Notomi et al, 2015;Tomita et al, 2008). As the LAMP technique uses strand-displacing DNA polymerases that have high activity under isothermal temperatures ranging from 60 to 68 uC, no expensive equipment is required (Iwamoto et al, 2003).…”

Section: Introductionmentioning

confidence: 99%

Detection of Mycobacterium tuberculosis complex in sputum specimens using a loop-mediated isothermal amplification assay in Korea

Moon

Kim

Tomono

et al. 2015

Journal of Medical Microbiology

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Tuberculosis (TB), an infectious disease caused by Mycobacterium tuberculosis complex (MTC), remains one of the leading causes of death in the world. In Korea, the current prevalence of multidrug-resistant TB (MDR-TB) poses a major problem. The most common method for diagnosing TB in developing countries is sputum smear microscopy; however, the sensitivity of this test is relatively low and it usually requires well-trained laboratory staff. Cultures of MTC require up to several weeks in sophisticated facilities, such as Biosafety Level 3. Effective diagnostic techniques are necessary to control TB. In Korea, we evaluated a loop-mediated isothermal amplification (LAMP) assay targeting the hspX gene (TB-hspX-LAMP) of MTC. For clinical evaluation, culture confirmation, smear microscopy and TB-hspX-LAMP were performed on 303 sputum specimens obtained from suspected TB patients in Korea. The sensitivity, specificity, positive predictive value and negative predictive value of TB-hspX-LAMP were 71.1, 98.8, 91.4 and 95.1 %, respectively, compared with TB culture, which is the gold standard for diagnosis of TB. In contrast, the comparable values of smear microscopy were 24.4, 98.1, 68.8 and 88.2 %, respectively. Therefore, we concluded that TB-hspX-LAMP was superior to the use of smear microscopy for the detection of MTC in sputum specimens in clinical settings in Korea. INTRODUCTIONTuberculosis (TB), an airborne infectious disease caused by Mycobacterium tuberculosis complex (MTC, Mycobacterium tuberculosis, Mycobacterium bovis, Mycobacterium africanum, Mycobacterium microti, Mycobacterium caprae, Mycobacterium canetti, Mycobacterium pinnipedii) (Behr et al., 1999; CDC, 2009), still remains one of the leading causes of death by infectious diseases worldwide. In 2013, 9 million people developed TB throughout the world; 1.5 million of this group died from it, of which 360 000 were human immunodeficiency virus-positive (WHO, 2014).Early, accurate diagnosis of MTC is important for the control of TB (Brodie & Schluger, 2005). The most common method for diagnosing TB in developing countries is sputum smear microscopy (George et al., 2011). Smear microscopy offers the advantages of rapid detection time and simplicity. However, the sensitivity of smear microscopy is relatively low and it requires well-trained laboratory staff.Culture methods can detect as few as 100 Mycobacterium cells per 1 ml specimen (Hofmann-Thiel et al., 2010) and TB culture is still considered the gold standard for the et al., 2003). The LAMP reaction produces stem-loop DNA structures with several inverted repeats and cauliflower-like structures with multiple loops. The products of amplified LAMP reactions can be measured by a change in the turbidity of the reaction mixture because magnesium pyrophosphate accumulates in the reaction mixture as a byproduct of the amplification reaction.The LAMP assay has several exceptional features compared with other TB diagnostic tools.(1) The LAMP assay uses four different primers to identify six distinct regions on...

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Loop-mediated isothermal amplification (LAMP): principle, features, and future prospects

Cited by 698 publications

References 14 publications

Rapid diagnosis of Zika virus through saliva and urine by Loop-mediated isothermal amplification (LAMP)

Rapid diagnosis of Zika virus through saliva and urine by Loop-mediated isothermal amplification (LAMP)

Loop‐mediated isothermal amplification (LAMP) as an alternative to PCR: A rapid on‐site detection of gene doping

Detection of Mycobacterium tuberculosis complex in sputum specimens using a loop-mediated isothermal amplification assay in Korea

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