scholarly journals Development of a portable reverse transcription loop‐mediated isothermal amplification system to detect the E1 region of Chikungunya virus in a cost‐effective manner

2020 ◽  
Author(s):  
Benjawan Saechue ◽  
Naganori Kamiyama ◽  
Yinan Wang ◽  
Chiaki Fukuda ◽  
Kei Watanabe ◽  
...  
Keyword(s):  
Reverse Transcription ◽  
Chikungunya Virus ◽  
Cost Effective ◽  
Isothermal Amplification ◽  
Loop Mediated Isothermal Amplification ◽  
Effective Manner ◽  
Amplification System

scholarly journals Portable and label-free quantitative Loop-mediated Isothermal Amplification (qLAMP) for reliable COVID-19 diagnostics in 3 minutes: An Arduino-based detection system assisted by a pH microelectrode

2021 ◽  
Author(s):  
Mario Moisés Alvarez ◽  
Sergio Bravo-González ◽  
Everardo González-González ◽  
Grissel Trujillo-de Santiago
Keyword(s):  
Real Time ◽  
Detection System ◽  
Genetic Material ◽  
Cost Effective ◽  
Isothermal Amplification ◽  
Diagnostic Methods ◽  
Label Free ◽  
Viral Pathogens ◽  
Loop Mediated Isothermal Amplification ◽  
Amplification System

Loop-mediated isothermal amplification (LAMP) has been recently studied as an alternative method for cost-effective diagnostics in the context of the current COVID-19 pandemic. Recent reports document that LAMP-based diagnostic methods have a comparable sensitivity and specificity to that of RT-qPCR. We report the use of a portable Arduino-based LAMP-based amplification system assisted by pH microelectrodes for the accurate and reliable diagnosis of SARS-CoV-2 during the first 3 minutes of the amplification reaction. We show that this simple system enables a straightforward discrimination between samples containing or not containing artificial SARS-CoV-2 genetic material in the range of 10 to 10,000 copies per 50 μL of reaction mix. We also spiked saliva samples with SARS-CoV-2 synthetic material and corroborated that the LAMP reaction can be successfully monitored in real time using microelectrodes in saliva samples as well. These results may have profound implications for the design of real-time and portable quantitative systems for the reliable detection of viral pathogens including SARS-CoV-2.

scholarly journals The detection of SARS-CoV-2 using reverse transcription loop-mediated isothermal amplification (RT-LAMP) in developing country

10.51511/pr.9 ◽  
2021 ◽  
Vol 1 (1) ◽  
pp. 9
Author(s):  
Muhammad Rizky Wibowo ◽  
Erna Harfiani ◽  
Sarmoko Sarmoko ◽  
Yudhi Nugraha
Keyword(s):  
Reverse Transcription ◽  
Rural Areas ◽  
Genetic Material ◽  
Cost Effective ◽  
Isothermal Amplification ◽  
World Health ◽  
Rt Pcr ◽  
Loop Mediated Isothermal Amplification ◽  
Alternative Detection ◽  
Health Organization

The severe acute respiratory syndrome coronavirus (SARS-CoV-2) has infected the human system resulting in Covid-19, and has spread rapidly worldwide. Therefore, a fast, simple, cost-effective, and accurate detecting tool is required. The standard diagnostic tool of the World Health Organization is the reverse transcription-polymerase chain reaction (RT-PCR). This method detects the presence of viral genetic material in the human body with accurate results. However, it has several limitations in terms of equipment, personnel, duration, and cost. Therefore, a fast, simple, and sensitive alternative detection, is required, one of which is the reverse transcription loop-mediated isothermal amplification (RT-LAMP) that functions under isothermal conditions. This method is battery-driven, hence, easy to move closer to the patient. Conclusively, the RT-LAMP test for SARS CoV-2 diagnosis produces comparable sensitivity to a standard RT-PCR and is more suitable for resource-poor settings, such as rural areas of developing countries.

scholarly journals Portable and Label-Free Quantitative Loop-Mediated Isothermal Amplification (LF-qLamp) for Reliable COVID-19 Diagnostics in Three Minutes of Reaction Time: Arduino-Based Detection System Assisted by a pH Microelectrode

Biosensors ◽  
2021 ◽  
Vol 11 (10) ◽  
pp. 386
Author(s):  
Mario Moisés Alvarez ◽  
Sergio Bravo-González ◽  
Everardo González-González ◽  
Grissel Trujillo-de Santiago
Keyword(s):  
Real Time ◽  
Detection System ◽  
Genetic Material ◽  
Cost Effective ◽  
Isothermal Amplification ◽  
Diagnostic Methods ◽  
Label Free ◽  
Viral Pathogens ◽  
Loop Mediated Isothermal Amplification ◽  
Amplification System

Loop-mediated isothermal amplification (LAMP) has been recently studied as an alternative method for cost-effective diagnostics in the context of the current COVID-19 pandemic. Recent reports document that LAMP-based diagnostic methods have a comparable sensitivity and specificity to that of RT-qPCR. We report the use of a portable Arduino-based LAMP-based amplification system assisted by pH microelectrodes for the accurate and reliable diagnosis of SARS-CoV-2 during the first 3 min of the amplification reaction. We show that this simple system enables a straightforward discrimination between samples containing or not containing artificial SARS-CoV-2 genetic material in the range of 10 to 10,000 copies per 50 µL of reaction mix. We also spiked saliva samples with SARS-CoV-2 synthetic material and corroborated that the LAMP reaction can be successfully monitored in real time using microelectrodes in saliva samples as well. These results may have profound implications for the design of real-time and portable quantitative systems for the reliable detection of viral pathogens including SARS-CoV-2.

scholarly journals Development and validation of a one-step reverse transcription loop-mediated isothermal amplification (RT-LAMP) for rapid detection of ZIKV in patient samples from Brazil

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Severino Jefferson Ribeiro da Silva ◽  
Keith Pardee ◽  
Udeni B. R. Balasuriya ◽  
Lindomar Pena
Keyword(s):  
Reverse Transcription ◽  
Rapid Detection ◽  
Quantitative Polymerase Chain Reaction ◽  
Rna Extraction ◽  
Lamp Assay ◽  
Isothermal Amplification ◽  
Serum Samples ◽  
Low Resource ◽  
Loop Mediated Isothermal Amplification ◽  
One Step

AbstractWe have previously developed and validated a one-step assay based on reverse transcription loop-mediated isothermal amplification (RT-LAMP) for rapid detection of the Zika virus (ZIKV) from mosquito samples. Patient diagnosis of ZIKV is currently carried out in centralized laboratories using the reverse transcription-quantitative polymerase chain reaction (RT-qPCR), which, while the gold standard molecular method, has several drawbacks for use in remote and low-resource settings, such as high cost and the need of specialized equipment. Point-of-care (POC) diagnostic platforms have the potential to overcome these limitations, especially in low-resource countries where ZIKV is endemic. With this in mind, here we optimized and validated our RT-LAMP assay for rapid detection of ZIKV from patient samples. We found that the assay detected ZIKV from diverse sample types (serum, urine, saliva, and semen) in as little as 20 min, without RNA extraction. The RT-LAMP assay was highly specific and up to 100 times more sensitive than RT-qPCR. We then validated the assay using 100 patient serum samples collected from suspected cases of arbovirus infection in the state of Pernambuco, which was at the epicenter of the last Zika epidemic. Analysis of the results, in comparison to RT-qPCR, found that the ZIKV RT-LAMP assay provided sensitivity of 100%, specificity of 93.75%, and an overall accuracy of 95.00%. Taken together, the RT-LAMP assay provides a straightforward and inexpensive alternative for the diagnosis of ZIKV from patients and has the potential to increase diagnostic capacity in ZIKV-affected areas, particularly in low and middle-income countries.

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