Loop-Mediated Isothermal Amplification Assay (LAMP): A Rapid Tool for Diagnosis of Foodborne and Zoonotic Pathogens: A Review

Zende, R. J.; Kshirsagar, D. P.; Vaidya, V. M.; Waghamare, R. N.; Todankar, R.P.; Shirke, A.H.

doi:10.5455/ijlr.20170415114515

Cited by 4 publications

(6 citation statements)

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“…The PCR and LAMP are the common methods used in the detection of the bacterium. These methods with the characteristics of high sensitivity and few steps have been applied to the detection of various pathogens, including fungi, parasites, viruses, and bacteria (31,32). However, they are still dependent on bacterium enrichment and genomic extraction, which would take several hours.…”

Section: Discussionmentioning

confidence: 99%

Visual and Rapid Detection of Klebsiella pneumoniae by Magnetic Immunocapture-Loop-Mediated Isothermal Amplification Assay

Zhang

Shi

Chen

et al. 2019

Jundishapur J Microbiol

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Background: Klebsiella pneumoniae is an important human pathogen that causes severe diseases including urinary tract infection, pneumonia, and bacteremia. However, a rapid and sensitive detection method remains to be developed. Objectives: This study aimed to develop a rapid, real-time, and visual detection method for K. pneumoniae. This is a primary screening method to improve the diagnosis of K. pneumoniae infection and save much precious time for clinical practice. Methods: Klebsiella pneumoniae was used as an antigen to produce a monoclonal antibodies (mAb). The mAb 1E6 recognizing outer membrane protein C on the surface of K. pneumoniae was screened by an indirect enzyme-linked immunosorbent assay (ELISA), Western Blot, and mass spectrometry assay, and then conjugated with the protein A/G coated magnetic beads to generate the immune magnetic beads (IMBs). Thereafter, the IMBs were used to capture K. pneumoniae, and the complex (beads-mAb-K. pneumoniae) was used for loop-mediated isothermal amplification (LAMP) assay. Results: We developed a rapid, real-time, and visual detection method employing an immunocapture loop-mediated isothermal amplification (IC-LAMP). The sensitivity of the IC-LAMP was 4 CFU mL-1. The process of K. pneumoniae detection lasted~60 minutes and had no cross-reaction to other microbial strains. Besides, the accuracy of IC-LAMP was further verified by examining 39 clinical isolates. Conclusions: Without the need for enrichment of bacteria and extraction of its genome, the IC-LAMP developed here could be used as a primary screening method supplementary to traditional detection methods to improve the diagnosis of K. pneumoniae infection for clinical practice.

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Section: Discussionmentioning

confidence: 99%

Visual and Rapid Detection of Klebsiella pneumoniae by Magnetic Immunocapture-Loop-Mediated Isothermal Amplification Assay

Zhang

Shi

Chen

et al. 2019

Jundishapur J Microbiol

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“…Além da Bst DNA polimerase e dos primers, a mistura reacional de LAMP consiste basicamente no uso de desoxinucleotídeos trifosfatos (dNTPs), sulfato de magnésio, betaína, tampão e DNA molde 99 .…”

Section: Lamp: Histórico E Descrição Da Técnicaunclassified

“…A LAMP tem inúmeras vantagens que a torna promissora como método de amplificação de ácido nucleico 20 . A primeira delas é que a técnica não requer equipamentos caros e devido a sua capacidade de amplificar o DNA sob condições isotérmicas, pode ser realizada em banho-maria, que é um equipamento simples e de baixo custo 68,75,99 .…”

Section: Vantagensunclassified

“…O subproduto da LAMP é um pirofosfato de magnésio que, por precipitar-se e gerar turbidez da solução, pode fornecer um indicador positivo da reação 99,65 . Embora quando o precipitado não for passível de visualização, haja a possibilidade de confirmar a presença do amplicon LAMP pela eletroforese em gel de agarose e/ou por sistemas colorimétricos que permitem a determinação do resultado a olho nu 65,66 .…”

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Loop Mediated Isothermal Amplification Assay (Lamp): Uma Revisão Detalhada Sobre a Técnica

Ribeiro

Cruz

2022

EVS

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A técnica de Loop Mediated Isothermal Amplification Assay (LAMP) amplamente difundida a partir dos anos 2000, baseia-se na utilização de primers sensíveis e específicos e na utilização da BST DNA polimerase junto à outros reagentes para amplificação de uma sequência alvo de DNA sob temperatura constante. Durante a LAMP ocorrem sucessivos ciclos de etapa não cíclica e cíclica para a produção exponencial de amplicons de comprimentos variáveis. Esses produtos da amplificação podem ser visualizados por eletroforese em gel de agarose, colorimetria e outras técnicas reveladoras da amplificação. Quanto à otimização da técnica, implica em planejamento da reação e execução de diversas procedimentos laboratoriais para confirmação dos resultados obtidos pela LAMP. De acordo com o objetivo do estudo do pesquisador, também é possível optar por uma das variações do ensaio LAMP, como a AS-LAMP, mLAMP e Id-LAMP. Assim, diante do exposto nesta pesquisa, o pesquisador e o público em geral podem conhecer mais sobre essa relevante técnica molecular que tende a passar por constatastes adaptações, reflexo nítido da evolução da ciência rumo à solução de questões cotidianas e à qualidade de vida da sociedade.

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“…In the past few decades, several molecular methods have been developed to overcome the shortcomings of the classical diagnostics methods, especially the in vitro amplification of a pathogen-specific nucleic acid sequence. Loop-mediated isothermal amplification technology developed by Notomi et al [10] is a novel DNA amplification method which can amplify target gene under isothermal conditions with high efficiency and sensitivity [24]. Developing countries like India require low-cost detection techniques for detection of these pathogens at district, block as well as at field level.…”

Section: Introductionmentioning

confidence: 99%

Rapid Detection of Shiga toxin-Producing E. Coli in Animal Origin Foods Using Loop-Mediated Isothermal Amplification (LAMP) Assay

et al. 2018

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The aim of this study was comparative evaluation of loop-mediated isothermal amplification (LAMP) and polymerase chain reaction (PCR) assay for rapid and inexpensive detection of shiga toxin-producing E. coli in animal origin foods by targeting stx1 and stx2 genes. The LAMP assay was performed using a water bath. The standardized LAMP assay was evaluated on 122 E. coli field isolates obtained from various animal origin food samples to ensure its reliability and usefulness. The result showed that conventional PCR could detect 68 (55.73%) and 75 (61.47%) positive E. coli isolates for stx1 and stx2 genes. Whereas, LAMP showed higher sensitivity by detecting 79 (64.75%) and 87 (71.31%) positive isolates of E. coli for stx1 and stx2 genes, respectively. LAMP assay was found to be highly specific and 10 times more sensitive as it could detect 1.11 9 10 2 cfu/ml for both stx1 and stx2 genes of E. coli isolates, whereas conventional PCR could detect 1.85 x 10 3 cfu/ml for both stx1 and stx2 genes of E. coli isolates. The rapidness, sensitivity, specificity, easiness and cost-effectiveness of LAMP assays will be very useful for the detection of foodborne pathogens for improving food sanitation and maintaining food safety.

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Loop-Mediated Isothermal Amplification Assay (LAMP): A Rapid Tool for Diagnosis of Foodborne and Zoonotic Pathogens: A Review

Abstract: Loop-mediated isothermal amplification (LAMP) assay was introduced in the year

Cited by 4 publications

References 5 publications

Visual and Rapid Detection of Klebsiella pneumoniae by Magnetic Immunocapture-Loop-Mediated Isothermal Amplification Assay

Visual and Rapid Detection of Klebsiella pneumoniae by Magnetic Immunocapture-Loop-Mediated Isothermal Amplification Assay

Loop Mediated Isothermal Amplification Assay (Lamp): Uma Revisão Detalhada Sobre a Técnica

Rapid Detection of Shiga toxin-Producing E. Coli in Animal Origin Foods Using Loop-Mediated Isothermal Amplification (LAMP) Assay

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