scholarly journals Precipitation of PEG/Carboxyl-Modified Gold Nanoparticles with Magnesium Pyrophosphate: A New Platform for Real-Time Monitoring of Loop-Mediated Isothermal Amplification

2017 ◽  
Vol 9 (12) ◽  
pp. 10472-10480 ◽  
Author(s):  
Ailin Qin ◽  
Lok Tin Fu ◽  
Jacky K. F. Wong ◽  
Li Yin Chau ◽  
Shea Ping Yip ◽  
...  
Keyword(s):  
Gold Nanoparticles ◽  
Real Time ◽  
Isothermal Amplification ◽  
Real Time Monitoring ◽  
Loop Mediated Isothermal Amplification ◽  
Magnesium Pyrophosphate

Screen-Printed Polyaniline-Based Electrodes for the Real-Time Monitoring of Loop-Mediated Isothermal Amplification Reactions

2017 ◽  
Vol 89 (19) ◽  
pp. 10124-10128 ◽  
Author(s):  
David Gosselin ◽  
Maxime Gougis ◽  
Mélissa Baque ◽  
Fabrice P. Navarro ◽  
Mohamed N. Belgacem ◽  
...  
Keyword(s):  
Real Time ◽  
Isothermal Amplification ◽  
Real Time Monitoring ◽  
The Real ◽  
Loop Mediated Isothermal Amplification ◽  
Screen Printed

scholarly journals Improved Visual Detection of speB Gene in Streptococcuspyogenes Isolates by Real-Time Loop-mediated Isothermal Amplification Turbidimetry Method

2021 ◽  
Vol 14 (2) ◽  
Author(s):  
Azi Simon Simon ◽  
Suresh Kumar Subbiah ◽  
Leslie Thian Lung Than ◽  
Malina Osman ◽  
Rukman Awang Hamat
Keyword(s):  
Real Time ◽  
Visual Detection ◽  
Clinical Manifestations ◽  
Lamp Assay ◽  
Healthcare Management ◽  
Isothermal Amplification ◽  
Real Time Monitoring ◽  
Gene Detection ◽  
The Real ◽  
Loop Mediated Isothermal Amplification

Background: Group A Streptococcus (GAS) causes a wide array of clinical manifestations ranging from mild pharyngitis to suppurative and non-suppurative severe debilitating diseases. Hence, a simple, rapid detection method with high sensitivity and specificity is needed. Objectives: This study embarked on the visual detection of the streptococcal pyrogenic exotoxin B (speB) gene by Real-time Turbidimetry and Loop-mediated Isothermal Amplification (RT-LAMP) methods. The real-time monitoring of the sigmoidal graph generated from a turbidometry method was incorporated in the assay. Methods: The amplification of the speB gene was virtually observed in real-time monitoring of the graph (sigmoidal curve) generated via a turbidimeter, thus providing a “guide” to accurately estimate the time to positivity for the gene detection. Results: The targeted gene was detected at 15 min but was optimally amplified within 45 min at an isothermal temperature of 63°C with 100% specificity using an established set of primers. The formation of sigmoidal curves was correlated with other visual observations by the naked eye (from orange to green), ultra-violet light (green fluorescence), and agarose gel electrophoresis. The improved detection limit of the real-time RT-LAMP assay was also observed compared to conventional PCR assay (0.001 pg/µL versus 1 ng/µL). Conclusions: In conclusion, this improved visual detection of RT-LAMP assay could provide additional insight for rapid, cost-effective, and reliable identification of GAS via speB gene detection in low or middle-income countries. It could also be a very important tool to improve the healthcare management of patients infected with GAS in the future.

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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