Loop Mediated Isothermal Amplification: A Promising Tool for Screening Genetic Mutations

Arjuna, Srividya; Maiti, Biswajit; Chakraborty, Anirban; Chakraborty, Gunimala

doi:10.1007/s40291-019-00422-0

Cited by 29 publications

(24 citation statements)

References 92 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Initially developed as a technique for rapid identification of infectious agents, LAMP has undergone extensive modifications in recent years 12 . AS-LAMP, which is an important variant of traditional LAMP, is now being appreciated as an effective alternative in SNP identification 11 . Previously, we developed AS-LAMP technique for screening germline and somatic de novo T790M mutation using blood and tissue DNA 13 .…”

Section: Discussionmentioning

confidence: 99%

“…LAMP assay relies on isothermal amplification of the desired gene fragment, using four primers, two outer (F3 and B3) and two inner (FIP and BIP) that mark six regions within the target sequence. The assay works on auto cycling strand displacement activity of target DNA by Bst polymerase, and results in the formation and accumulation of stem loop DNA 11 . In this study, we have developed novel, cell free DNA-based allele specific loop-mediated isothermal amplification (CF-LAMP) assay for screening two clinically relevant mutations, T790M (C>T) point mutation in exon 20 and L858R (T>G) point mutation in exon 21 of EGFR gene.…”

mentioning

confidence: 99%

See 1 more Smart Citation

Non-invasive detection of EGFR mutations by cell-free loop-mediated isothermal amplification (CF-LAMP)

Arjuna

Venkataram

Dechamma

et al. 2020

Sci Rep

Self Cite

View full text Add to dashboard Cite

Targeting epidermal growth factor receptor (EGFR) through tyrosine kinase inhibitors (TKI) is a successful therapeutic strategy in non-small cell lung cancer. However, the response to TKI therapy depends on specific activating and acquired mutations in the tyrosine kinase domain of the EGFR gene. Therefore, confirming the EGFR status of patients is crucial, not only for determining the eligibility, but also for monitoring the emergence of mutations in patients under TKI therapy. In this study, our aim was to develop a cost effective, yet sensitive, technique that allows the detection of therapeutically-relevant EGFR hotspot mutations at isothermal conditions in a non-invasive manner. Previously, we developed an allele-specific loop-mediated isothermal amplification (AS-LAMP) assay for screening germline and somatic de novo T790M EGFR mutation in lung cancer patients. In this study, we used cell free DNA as a template in AS-LAMP assay (CF-LAMP) for non-invasive detection of two hotspot EGFR mutations (T790M, and L858R) and compared its efficiency with ultrasensitive droplet digital PCR (ddPCR) assay. The results of CF-LAMP assay were consistent with those obtained in ddPCR assay, indicating the robustness of the method. CF-LAMP may serve as a valuable and cost-effective alternative for liquid biopsy techniques used in molecular diagnosis of non-small cell lung cancer.

show abstract

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

Non-invasive detection of EGFR mutations by cell-free loop-mediated isothermal amplification (CF-LAMP)

Arjuna

Venkataram

Dechamma

et al. 2020

Sci Rep

Self Cite

View full text Add to dashboard Cite

show abstract

“…[6][7][8][9] LAMP assays are also an effective tool for screening genetic mutations. 10 A previous study detected the -SEA (Southeast Asian) deletion, and IVSII-I G>A mutation indicating thalassemia using LAMP assays. 11,12 Thus, new LAMP methods capable of detecting thalassemia could be alternative methods to other techniques, and could be used in primary hospitals and rural areas.…”

Section: Introductionmentioning

confidence: 99%

<p>Establishment and Evaluation of a Novel Method Based on Loop-Mediated Isothermal Amplification for the Rapid Diagnosis of Thalassemia Genes</p>

Lin¹,

Li²,

Huang³

et al. 2020

RMHP

View full text Add to dashboard Cite

Currently, thalassemia is commonly detected using gap-polymerase chain reaction (PCR) and deoxyribonucleic acid (DNA) reverse dot blot, which have high requirements of space, instruments, and personnel. Therefore, it is necessary to develop a new method for thalassemia detection with high sensitivity, low cost, and simple and fast operation. In this study, we aimed to design and evaluate a new method for detecting three α-thalassemia genes including-Southeast Asian (SEA),-α3.7, and-α4.2 and five β-thalassemia genes including 654M, 41/42M, −28M, 17M, and 27/28M based on loop-mediated isothermal amplification (LAMP). Methods: Primer sequences were designed using Primer Explorer V4 software. Blood samples (5 mL) were collected from all participants in EDTA. DNA was extracted using Chelex 100 and was subjected to LAMP. LAMP products were detected by fluorescence development in ultraviolet light. Results: We found that LAMP assays for positive samples of thalassemia reached a plateau before 60 minutes, whereas the negative control samples entered the plateau after 70 minutes or showed no amplification. The concentration range of positive reactions was between 20-60 pg/μL and 20-60 ng/μL. Additionally, there were no cross-reactivities among 8 thalassemia subtypes. For clinical samples, the positive sample tube showed strong green fluorescence, whereas the negative tube showed light green fluorescence. According to these results, the LAMP method has high sensitivity for detecting thalassemia (252/254). However, 43 false-positive results were obtained in the LAMP test. The LAMP assay was also of low cost and with simple and fast operation. Conclusion: The novel LAMP assay can be completed within 60 min using a heating block or a water bath, and the result can be read visually based on color change to detect thalassemia. The LAMP assay fulfills the requirements of field application and resourcelimited areas, especially those with primary hospitals and rural areas.

show abstract

“…Since this first report, the application of LAMP has been increasingly used and adopted as an alternative method to those based on PCR. LAMP, in fact, is continuously being implemented in the medicine, agriculture, and food industries, with approaches that include the screening of viral and bacterial strain mutations, analysis of fungicide resistant mutations, analysis of micro RNAs, herbal medicine identification, plant pathogen vectors identification, single nucleotide polymorphisms analysis, and detection of genetically modified organisms [16][17][18][19][20]. The reasons for the development of the LAMP methodology were founded on the attempt to overcome some drawbacks of the conventional PCR, a method that requires the acquisition of a high-cost equipment known as thermal cycler.…”

Section: Loop Mediated Isothermal Amplification (Lamp)mentioning

confidence: 99%

Loop Mediated Isothermal Amplification: Principles and Applications in Plant Virology

Panno

Matić

Tiberini³

et al. 2020

Plants

140

111

View full text Add to dashboard Cite

In the last decades, the evolution of molecular diagnosis methods has generated different advanced tools, like loop-mediated isothermal amplification (LAMP). Currently, it is a well-established technique, applied in different fields, such as the medicine, agriculture, and food industries, owing to its simplicity, specificity, rapidity, and low-cost efforts. LAMP is a nucleic acid amplification under isothermal conditions, which is highly compatible with point-of-care (POC) analysis and has the potential to improve the diagnosis in plant protection. The great advantages of LAMP have led to several upgrades in order to implement the technique. In this review, the authors provide an overview reporting in detail the different LAMP steps, focusing on designing and main characteristics of the primer set, different methods of result visualization, evolution and different application fields, reporting in detail LAMP application in plant virology, and the main advantages of the use of this technique.

show abstract

Loop Mediated Isothermal Amplification: A Promising Tool for Screening Genetic Mutations

Cited by 29 publications

References 92 publications

Non-invasive detection of EGFR mutations by cell-free loop-mediated isothermal amplification (CF-LAMP)

Non-invasive detection of EGFR mutations by cell-free loop-mediated isothermal amplification (CF-LAMP)

<p>Establishment and Evaluation of a Novel Method Based on Loop-Mediated Isothermal Amplification for the Rapid Diagnosis of Thalassemia Genes</p>

Loop Mediated Isothermal Amplification: Principles and Applications in Plant Virology

Contact Info

Product

Resources

About