scholarly journals Non‐lethal loop‐mediated isothermal amplification assay as a point‐of‐care diagnostics tool for Neoparamoeba perurans , the causative agent of amoebic gill disease

2020 ◽  
Vol 43 (7) ◽  
pp. 779-790 ◽  
Author(s):  
Irene Cano ◽  
Robin McCullough ◽  
Brian Mulhearn ◽  
Susie Gunning ◽  
Ava Waine ◽  
...  
Keyword(s):  
Causative Agent ◽  
Point Of Care ◽  
Isothermal Amplification ◽  
Loop Mediated Isothermal Amplification ◽  
Point Of Care Diagnostics ◽  
Amplification Assay ◽  
Gill Disease

scholarly journals Progress in loop-mediated isothermal amplification assay for detection of Schistosoma mansoni DNA: towards a ready-to-use test

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
J. García-Bernalt Diego ◽  
P. Fernández-Soto ◽  
B. Crego-Vicente ◽  
S. Alonso-Castrillejo ◽  
B. Febrer-Sendra ◽  
...  
Keyword(s):  
Schistosoma Mansoni ◽  
Molecular Diagnostics ◽  
Neglected Tropical Diseases ◽  
Point Of Care ◽  
Isothermal Amplification ◽  
Diagnostic Tools ◽  
Important Species ◽  
Loop Mediated Isothermal Amplification ◽  
Amplification Assay ◽  
Diagnostic Applications

Abstract Schistosomiasis is one of the most prevalent Neglected Tropical Disease, affecting approximately 250 million people worldwide. Schistosoma mansoni is the most important species causing human intestinal schistosomiasis. Despite significant efforts in recent decades, the global disease burden of schistosomiasis remains extremely high. This could partly be attributed to the absence of accurate diagnostic tools, primarily in endemic areas. Loop-mediated isothermal amplification (LAMP) is increasingly used in molecular diagnostics as a field-friendly alternative to many other complex molecular methods and it has been proposed as an ideal candidate for revolutionizing point-of-care molecular diagnostics. In a previous work, a LAMP-based method to detect S. mansoni DNA (SmMIT-LAMP) was developed by our research group for early diagnosis of active schistosomiasis in an experimental infection murine model. The SmMIT-LAMP has been further successfully evaluated in both human stool and snail samples and, recently, in human urine samples. In this study, we developed an important improvement for SmMIT-LAMP molecular assay, transforming it into a cold maintenance dry format suitable for potentially manufacturing as kit for ready-to-use for schistosomiasis diagnosis. This procedure could be applied to create dry LAMP kits for a laboratory setting and for diagnostic applications for other neglected tropical diseases.

scholarly journals Rapid and sensitive detection of Salmonella species targeting the hilA gene using a loop-mediated isothermal amplification assay

2021 ◽  
Vol 19 (3) ◽  
pp. e30
Author(s):  
Jiyon Chu ◽  
Juyoun Shin ◽  
Shinseok Kang ◽  
Sun Shin ◽  
Yeun-Jun Chung
Keyword(s):  
Point Of Care ◽  
Lamp Assay ◽  
Conventional Polymerase Chain Reaction ◽  
Isothermal Amplification ◽  
Sensitive Detection ◽  
Conventional Pcr ◽  
Pcr Assay ◽  
Loop Mediated Isothermal Amplification ◽  
Amplification Assay ◽  
Specific Amplification

Salmonella species are among the major pathogens that cause foodborne illness outbreaks. In this study, we aimed to develop a loop-mediated isothermal amplification (LAMP) assay for the rapid and sensitive detection of Salmonella species. We designed LAMP primers targeting the hilA gene as a universal marker of Salmonella species. A total of seven Salmonella species strains and 11 non-Salmonella pathogen strains from eight different genera were used in this study. All Salmonella strains showed positive amplification signals with the Salmonella LAMP assay; however, there was no non-specific amplification signal for the non-Salmonella strains. The detection limit was 100 femtograms (20 copies per reaction), which was ~1,000 times more sensitive than the detection limits of the conventional polymerase chain reaction (PCR) assay (100 pg). The reaction time for a positive amplification signal was less than 20 minutes, which was less than one-third the time taken while using conventional PCR. In conclusion, our Salmonella LAMP assay accurately detected Salmonella species with a higher degree of sensitivity and greater rapidity than the conventional PCR assay, and it may be suitable for point-of-care testing in the field.

DETECTION METHODS FOR RESULTS OF A LOOP-MEDIATED ISOTHERMAL AMPLIFICATION OF DNA

2020 ◽  
Vol 65 (1) ◽  
pp. 67-72
Author(s):  
Olga A. Petrusha ◽  
E. B. Faizuloev
Keyword(s):  
Gold Nanoparticles ◽  
Real Time ◽  
Fluorescence Detection ◽  
Point Of Care ◽  
Colorimetric Detection ◽  
Lamp Reaction ◽  
Isothermal Amplification ◽  
Loop Mediated Isothermal Amplification ◽  
Advantages And Disadvantages ◽  
Point Of Care Diagnostics

The loop mediated isothermal amplification (LAMP) was developed by T. Notomi et al. in 2000. It has become one of the most promising methods for point-of-care diagnostics due to its accuracy, sensitivity and ease of execution. In this review, various methods for detecting the results of the LAMP reaction are considered; their advantages and disadvantages are revealed. Methods for detecting LAMP results can be divided into indirect and direct. Indirect methods aimed at detecting changes in the chemical composition of the reaction mixture include real-time turbidimetry, fluorescence detection with calcein, colorimetric detection with hydroxynaphthol blue, and detection using modified gold nanoparticles. Direct methods based on the detection of accumulation amplicons during the reaction include fluorimetric detection with intercalating dyes, resonance fluorescence energy transfer, enzyme immunoassay, immunochromatography, using cationic polymers and gold nanoparticles. The development in the field of point-of-care diagnostics is characterized by a pronounced tendency to miniaturization, the LAMP reaction on microchips and microfluidic devices with an electrochemical or optical detection method. The most promising for the diagnosis of infectious diseases are turbidimetry methods and the use of intercalating dyes. The development of portable domestic instruments for detecting of LAMP results based on real-time fluorescence detection or turbidimetry will contribute to the widespread introduction of the method into clinical laboratory diagnostic practice. A literature research was conducted in the Pubmed ncbi based on keywords.

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