Method for detecting genetic material after an amplification reaction after a loop-mediated isothermal amplification reaction.
Colorimetric method based on a complexometric indicator, for detecting genetic material after an amplification reaction after a loop-mediated isothermal amplification reaction
Loop-mediated isothermal amplification (LAMP) is a DNA amplification method that allows rapid and sensitive detection of a specific gene. LAMP merged with reverse transcription (RT-LAMP) has been successfully used for the detection of several respiratory RNA viruses, including SARS-CoV-2. RT-LAMP is a powerful alternative to RT-PCR due to its high specificity and sensitivity, cost effectiveness, and fast turnaround. The simple technical and instrumental requirements of colorimetric RT-LAMP tests make them extremely attractive for point-of-care (POC) use and implementation in low-resource settings.
Colorimetric RT-LAMP has been successfully used for detection of SARS-CoV-2in COVID-19 patients.
The colorimetric method for the detection of an amplified genetic material in a biological sample comprises the following steps:
- obtaining a biological sample
- amplifying the genetic material to obtain a solution of amplified genetic material
- adding a detection ensemble comprising murexide and a zinc (ll) compound to the solution of amplified genetic material
- evaluating the color change of the solution.
An aspect of the present disclosure also relates to the use of murexide indicator (5,5-Nitrilodibarbituric acid monoammonium salt, or ammonium purpurate, CAS 3051-09-0) and a zinc (ll) compound for detection of an amplified genetic material in a biological sample. A kit, a microfluidic diagnostic cartridge or a lab-on-chip device for detection of an amplified genetic material in a biological sample by the described method is also available. The method is also suitable for the detection of viral genetic material, bacterial genetic material, fungal genetic material, protozoal parasite genetic material or human genetic material.
STAGE OF DEVELOPMENT
Technology Readiness Level (TRL): 3 – Experimental Proof of Concept
BENEFITS & APPLICATIONS
Saliva-based testing has numerous advantages over nasopharyngeal sampling, especially in a mass screening scenario. It can be performed easily and non-invasively, thus minimizing patient discomfort, and it does not require specialized personnel or the use of protective equipment, which saves time and reduces costs.
Easiness of use is translated by the use of a kit, a microfluidic diagnostic cartridge or a lab-on-chip device for detection of an amplified genetic material in a biological sample by the described method.
- Available for exclusive licensing and/or seeking co-development partners.
Luís Miguel Lima