FRET-Based Assimilating Probe for Sequence-Specific Real-Time Monitoring of Loop-Mediated Isothermal Amplification (LAMP)

2011 ◽  
Vol 4 (2) ◽  
pp. 81-100 ◽  
Author(s):  
R. Kubota ◽  
A. M. Alvarez ◽  
W.W. Su ◽  
D. M. Jenkins
Keyword(s):  
Real Time ◽  
Isothermal Amplification ◽  
Real Time Monitoring ◽  
Loop Mediated Isothermal Amplification

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.

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