Loop-mediated isothermal amplification

LAMP analysis of wastewater (Anal. Chem. 2017)

Loop-mediated isothermal amplification (LAMP) is a single-tube technique for the amplification of DNA. It is a simple, rapid, and cost-effective method that has been used in the detection of various genetic diseases, viral, bacterial, and parasitic infections. Unlike polymerase chain reaction (PCR), LAMP carries out DNA amplification at a constant temperature, using a set of four specially designed primers that recognize a total of six distinct regions on the target DNA. This unique feature of LAMP allows for high specificity and efficiency in the amplification process.

Principles of LAMP

LAMP operates under isothermal conditions, typically ranging from 60 to 65 degrees Celsius. The process involves the use of a DNA polymerase with high strand displacement activity in addition to the set of four primers. The amplification cycle consists of the initial step of synthesizing a DNA loop, followed by cycling amplification steps that lead to the accumulation of amplification products with various structures, including stem-loop DNAs with several inverted repeats of the target and cauliflower-like structures with multiple loops.

Advantages of LAMP

The main advantages of LAMP over traditional PCR include:

• Isothermal amplification, eliminating the need for sophisticated thermal cycling equipment.
• High specificity due to the use of four to six primers recognizing six to eight regions on the target DNA.
• Rapid amplification, with results typically available within an hour.
• The possibility to visually detect the amplification products, either by turbidity or by fluorescence under UV light when using intercalating dyes.

Applications of LAMP

LAMP has been applied in various fields including:

• Clinical diagnostics, for the detection of pathogenic organisms and genetic diseases.
• Food safety, for the identification of foodborne pathogens.
• Environmental monitoring, for the detection of microbial contaminants.
• Agricultural research, for the identification of plant diseases and pests.

Limitations

While LAMP offers several advantages, it also has limitations:

• The risk of carryover contamination due to the high amounts of DNA produced.
• The complexity of primer design, which requires a good understanding of the target DNA regions.
• Potential for false positive results, necessitating careful interpretation of results.

Conclusion

LAMP is a versatile and powerful tool for the amplification of DNA under isothermal conditions. Its simplicity, efficiency, and cost-effectiveness make it suitable for a wide range of applications, particularly in settings with limited resources. Despite its limitations, ongoing research and development are likely to expand its utility and address current challenges.

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