RT-LAMP Assay

Loop-Mediated Isothermal Amplification Assay




LAMP Visual diagnostic test is a colorimetric RT (reverse transcriptase)- LAMP test intended for detection of nucleic acids from the SARS-CoV-2 virus.


SARS-CoV-2 RNA can be found in liquid from upper or lower respiratory tracts of infected individuals. Samples can be obtained by nasopharyngeal or oropharyngeal swabs or from sputum.


LAMP Visual can be analyzed by either real-time PCR signal or colorimetric observation by the naked eye. Positive LAMP results indicate an infection with SARS-CoV-2 virus.  As infection with SARS-CoV-2 can occur without showing any symptoms, negative LAMP results do not exclude present or hinder future infection with SARS-CoV-2 virus and the result should always be combined with clinical observations, patient history, and epidemiological information.


The test is applicable on standard real-time PCR equipment using standard procedures or in a simple heating block. The tubes contain synthetic DNA analogues comprising a flat heteroaromatic, hydrophobic molecule and a linker. These modifications are inserted into the oligonucleotides at fixed positions during synthesis.


 

Advantage of the test:


Fast: the results are ready in 30 minutes


Very high sensitivity: The RT-Lamp assay is basically as sensitive is RT-PCR test


Highly specific (SARS-CoV-2 Virus Specific): LAMP targets one marker, N1, of the SARS-CoV-2 nucleocapsid protein gene.


Easy to evaluate: Detection can be simply done by naked eye by checking the change of the color in the tubes.


Massive sample handling is possible: Multiple samples can be analyzed at the same time.  


Easy to use: No complicated laboratory equipment is needed. The test can be done on simple heating block on 65 degrees siliceous.


Minimum hands on handling: No pre-treatment of the sample is needed.  The test is ready to be used Immediately.


Easy to work: Minimal training is needed. However due to the contamination risk by COVID-19 virus it is recommended to be performed by trained specialists.

 


The technology behind:


Loop-mediated isothermal amplification (LAMP) is an isothermal amplification method where DNA/RNA can be amplified at a single temperature and allows colorimetric detection of the amplified product, eliminating the need for sophisticated instrumentation. The LAMP method utilizes 6 primers and a polymerase with strand displacement activity for fast and specific amplification.


Our LAMP assay is supplied as a Ready-To-Use (RTU) version. It is pre-dispensed in two tubes and needs only addition of RNA before running. The control assay is marked with a red dot for the ease of recognition.



In control tube, the detection of amplified human RNase P is used to determine whether a sample is valid buy testing the presence of nucleic acids in the sample. Since the RNase P assay can amplify both RNA and genomic DNA it acts only as a purification control. RNase P assays are marked with a red dot on the side of the PCR tube.

 


 

Colorimetric detection:


An overview of the possible outcomes of the analysis is shown below. The results are only valid if the included positive control has turned yellow post amplification in both RNase P and N1 tube. No template (NTC) negative control should remain pink after reaction.