Novel Molecular Diagnostic Platform for Tropical Infectious Diseases

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

doi:10.5772/29561

Cited by 5 publications

(4 citation statements)

References 26 publications

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“…Recently, a rapid and simplified molecular technique, the loop-mediated isothermal amplification (LAMP) has been shown to be an effective tool in detection of human pathogenic infectious agents (Notomi et al, 2000;Mori et al, 2012;Dhama et al, 2014). The technique has been applied in the detection of Leishmania using purified DNA extracted from patient's materials (Takagi et al, 2009;Adams et al, 2010;Khan et al, 2012) or swab boiled samples from CL model mice (Direct Boil-LAMP method; Mikita et al, 2014).…”

Section: Introductionmentioning

confidence: 99%

A rapid molecular diagnosis of cutaneous leishmaniasis by colorimetric malachite green-loop-mediated isothermal amplification (LAMP) combined with an FTA card as a direct sampling tool

et al. 2016

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Leishmaniasis remains one of the world's most neglected diseases, and early detection of the infectious agent, especially in developing countries, will require a simple and rapid test. In this study, we established a quick, one-step, single-tube, highly sensitive loop-mediated isothermal amplification (LAMP) assay for rapid detection of Leishmania DNA from tissue materials spotted on an FTA card. An FTA-LAMP with pre-added malachite green was performed at 64 degrees C for 60 mm using a heating block and/or water bath and DNA amplification was detected immediately after incubation. The LAMP assay had high detection sensitivity down to a level of 0.01 parasites per mu l. The field- and clinic-applicability of the colorimetric FTA-LAMP assay was demonstrated with 122 clinical samples collected from patients suspected of having cutaneous leishmaniasis in Peru, from which 71 positives were detected. The LAMP assay in combination with an FTA card described here is rapid and sensitive, as well as simple to perform, and has great potential usefulness for diagnosis and surveillance of leishmaniasis in endemic areas. (C) 2015 Elsevier B.V. All rights reserved

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Section: Introductionmentioning

confidence: 99%

A rapid molecular diagnosis of cutaneous leishmaniasis by colorimetric malachite green-loop-mediated isothermal amplification (LAMP) combined with an FTA card as a direct sampling tool

et al. 2016

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“…Specifically, LAMP employs two inner primers (FIP and BIP, which in turn consist of two parts each) and two outer primers (F3 and B3 which can recognize a total of six distinct regions within the target DNA). Two loop primers are employed (Forward loop primer; LF, and backward loop primer; LB) to accelerate amplification and detection efficiency [21][22][23].…”

Section: Evaluation Of the Hypothesis: Lamp-based Methods Of Viral Dementioning

confidence: 99%

“…These reactions result in a product with a dumbbell-like structure which is essential for LAMP to establish isothermal amplification as the loop structures are always single stranded and can be annealed by FIP or BIP. This loop structure formation eliminates the denaturing step, which is otherwise essential in PCR for obtaining single-stranded DNA, and also establishes a cyclic reaction between the dumbbell-like structure and its complementary product, leading to elongated products with various copies of the target sequence produced (for reviews and detailed schematics, refer to [22,23]). Numerous studies have now shown the successful application of LAMP assays in various forms to detect coronavirus RNA in patients samples [24][25][26][27], demonstrating that 1-10 copies of viral RNA template per reaction was sufficient for successful detection, which were~100fold more sensitive than conventional RT-PCR methods [26][27][28][29][30][31].…”

Section: Evaluation Of the Hypothesis: Lamp-based Methods Of Viral Dementioning

confidence: 99%

Loop mediated isothermal amplification (LAMP) assays as a rapid diagnostic for COVID-19

2020

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Recently, a novel coronavirus (SARS-CoV-2; coronavirus disease 2019, COVID-19) has emerged, rapidly spreading and severely straining the capacity of the global health community. Many nations are employing combinations of containment and mitigation strategies, where early diagnosis of COVID-19 is vital in controlling illness progression and limiting viral spread within the population. Thus, rapid and accurate methods of early detection are vital to contain COVID-19 and prevent further spread and predicted subsequent infectious waves of viral recurrence in future. Immediately after its initial characterization, Chinese and American Centers for Disease Control and Prevention (CDCs) rapidly employed molecular assays for detection of COVID-19, mostly employing real-time polymerase chain reaction (RT-PCR) methods. However, such methods require specific expensive items of equipment and highly trained analysts, requiring upwards of 4-8 h to process. These requirements coupled with associated financial pressures may prevent effective deployment of such diagnostic tests. Loop mediated isothermal amplification (LAMP) is method of nucleic acid amplification which exhibits increased sensitivity and specificity are significantly rapid, and do not require expensive reagents or instruments, which aids in cost reduction for coronavirus detection. Studies have shown the successful application of LAMP assays in various forms to detect coronavirus RNA in patient samples, demonstrating that 1-10 copies of viral RNA template per reaction are sufficient for successful detection,~100-fold more sensitive than conventional RT-PCR methods. Importantly, studies have also now demonstrated the effectiveness of LAMP methodology in the detection of SARS-CoV-2 RNA at significantly low levels, particularly following numerous improvements to LAMP assay protocols. We hypothesise that recent advancements in enhanced LAMP protocols assay perhaps represent the best chance for a rapid and robust assay for field diagnosis of COVID-19, without the requirement of specialized equipment and highly trained professionals to interpret results. Herein, we present our arguments with a view to disseminate such findings, to assist the combat of this virus that is proving so devastating. We hope that this strategy could be applied rapidly, and confirmed for viability with clinical samples, before being rolled out for mass-diagnostic testing in these current times.

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“…[82] For ASTs, LAMP is used in the determination of genes that could indicate the presence of resistance strains. [83][84][85] With this focus, Takano and collaborators [86] developed LAMP methods that detected genes related to antibiotic resistance in P. aeruginosa, GES (Guiana extended-spectrum) β-lactamase genes and carbapenemase genotypes. Gong et al [87] also reported the development of a LAMP amplification assay combined with a nanoparticle-based lateral flow biosensor (LFB) for the detection of a gene that confers resistance against colistin.…”

Section: Genomic Methodsmentioning

confidence: 99%

Antibiotic Resistance Detection in Pseudomonas aeruginosa: Recent Strategies, Advances, and Challenges

Clarindo Lopes,

Koushanpour,

Wiebe

et al. 2024

Analysis & Sensing

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Antibiotic resistance presents a worldwide public health emergency, impeding the effective management of infectious diseases. Pseudomonas aeruginosa plays a substantial role as a bacterial pathogen, particularly in infections among hospitalized individuals, and those with weakened immune systems. Timely and accurate detection of antimicrobial resistance in P. aeruginosa is crucial for initiating tailored antibiotic therapy promptly, thus improving patient outcomes. Nevertheless, this endeavor encounters challenges due to the intricate and varied nature of antibiotic‐resistant strains. Extensive efforts have been invested in developing sensors and instrumentations for assessing antibiotic resistance or susceptibility (AST), aiming to enable personalized patient treatment with appropriate medications. This paper focuses on recent advancements in these methodologies, utilized for evaluating the degree of antibiotic resistance or susceptibility inpathogenic bacteria. Flow cytometry, dual molecular recognition and mass spectrometry are presented as newer phenotypic AST techniques. Genomic methods and the pyocyanin detection are listed as methodologies for the detection of resistance indicators.

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Novel Molecular Diagnostic Platform for Tropical Infectious Diseases

Cited by 5 publications

References 26 publications

A rapid molecular diagnosis of cutaneous leishmaniasis by colorimetric malachite green-loop-mediated isothermal amplification (LAMP) combined with an FTA card as a direct sampling tool

A rapid molecular diagnosis of cutaneous leishmaniasis by colorimetric malachite green-loop-mediated isothermal amplification (LAMP) combined with an FTA card as a direct sampling tool

Loop mediated isothermal amplification (LAMP) assays as a rapid diagnostic for COVID-19

Antibiotic Resistance Detection in Pseudomonas aeruginosa: Recent Strategies, Advances, and Challenges

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