Language
Country/Area
No result found
How to use LAMP technology to perform DNA detection without a thermal cycler?
In today's biomedical field, the demand for DNA testing is increasing day by day, especially after the epidemic, public health has become extremely important. Although traditional polymerase chain reaction (PCR) is currently the most commonly used nucleic acid amplification technology, it relies on high-cost thermal cyclers, which limits its application in low-resource environments. At this time, Loop-mediated isothermal amplification (LAMP) technology is particularly important. It is a technology that performs nucleic acid amplification at a constant temperature, avoiding the complexity and equipment requirements of traditional methods.
LAMP was first developed in 1998 by Japan's Eiken Chemical Company to provide a low-cost and efficient alternative for DNA detection.
In the LAMP process, the target sequence is usually amplified at a constant temperature of 60 to 65°C. Unlike PCR technology, which requires multiple temperature changes, LAMP can continue to react at a single temperature. This technique typically uses four different primers to amplify six specific sites on the target gene, and its high specificity makes it ideal for many diagnostic applications.
Flexibility of detection methods
In terms of detection, LAMP products can be detected by a photometer, using the turbidity changes caused by inorganic salt precipitation (such as magnesium tetraphosphate) produced during the amplification process. This change can be easily identified with the naked eye or measured with a simple photometer. In addition, LAMP technology can also be used for point-of-care detection using intercalating dyes such as SYTO 9, which change fluorescence intensity as DNA amplifies.
For example, when gold nanoparticle-bound single-stranded DNA is used, the LAMP product can be visually detected by binding to its complementary DNA. This makes the LAMP method useful in reducing detection time and confirming the specificity of the amplified product. has advantages.
Benefits of using LAMP technology
The birth of LAMP provides a simple choice for clinical sites and testing. Because it does not require expensive thermal cyclers, LAMP is particularly suitable for the diagnosis of infectious diseases in countries with limited resources. In recent years, LAMP has been widely used in the detection of a variety of infectious diseases, such as malaria, tuberculosis, and SARS-CoV-2, and has shown its potential in developing countries.
In addition, LAMP shows higher resistance to inhibitors in complex samples (such as blood), which makes its application in the diagnostic field very valuable.
The simplicity of the technology also allows researchers to use a variety of indicators for easier analysis of results, such as detecting DNA through color changes, in fact making it easy to analyze test results with smartphones.
Limitations of LAMP technology
Although LAMP has many advantages, its limitations cannot be ignored. First, LAMP is less diverse than PCR and is mainly used for diagnosis or detection, but is not suitable for cloning or other molecular biology applications. In addition, since the primer design of LAMP is subject to various constraints, it becomes relatively more difficult to "manually design" primers. Although there are various software to assist in design, compared with PCR, its flexibility is still lacking.
Furthermore, during the late amplification process of LAMP, the interaction between primers may lead to non-specific amplification, which is one of the main factors affecting the accuracy of the results. This reinforces the need for precise sample handling and reaction conditions.
Future research directions
Recent research points out that efforts to combine LAMP technology with other reporting methods are gradually increasing. Among them, RNase hybrid-assisted amplification (RHAM) is a new attempt to combine LAMP with fluorescent reporting technology. This shows the potential for further development of LAMP technology in the future, which is expected to address existing limitations.
With the development of biomedical technology, LAMP technology has shown great potential in improving diagnostic effectiveness and accessibility. Can the widespread application of this technology change the way we face disease diagnosis?
