LAMP Method as One of the Best Candidates for Replacing with PCR Method
Journal name: The Malaysian Journal of Medical Sciences
Original article title: LAMP Method as One of the Best Candidates for Replacing with PCR Method
The Malaysian Journal of Medical Sciences (MJMS) is a peer-reviewed, open-access journal published online at least six times a year. It covers all aspects of medical sciences and prioritizes high-quality research.
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Masoud Keikha
The Malaysian Journal of Medical Sciences:
(A peer-reviewed, open-access journal)
Full text available for: LAMP Method as One of the Best Candidates for Replacing with PCR Method
Year: 2018 | Doi: 10.21315/mjms2018.25.1.15
Copyright (license): CC BY 4.0
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Summary of article contents:
Introduction
Molecular methods have revolutionized the field of microbiology, with the polymerase chain reaction (PCR) being one of the most significant advancements. PCR is revered for its rapid and sensitive ability to amplify minimal quantities of target DNA by several million-fold, making it a pivotal technique in medical microbiology. However, it does come with limitations such as the need for thermal cycling, specialized equipment, and toxic detection reagents like Ethidium bromide. These constraints often confine its usage to well-equipped laboratories managed by skilled personnel, highlighting the urgent need for simpler, more accessible methods.
Loop Mediated Isothermal Amplification (LAMP)
One promising alternative to PCR is Loop Mediated Isothermal Amplification (LAMP), a technique developed by Notomi et al. in 2000. LAMP stands out due to its isothermal amplification process that does not require thermal cycling apparatus like a thermocycler. It uses multiple primers to enhance specificity and sensitivity, allowing for amplification at a constant temperature of 59 °C to 66 °C. The process yields a stem-loop DNA structure, enhancing the detection accuracy through visual indicators, such as turbidity or the use of colorimetric dyes. LAMP has been successfully employed in diagnosing various infectious diseases, including those caused by bacteria such as Mycobacterium tuberculosis, Staphylococcus aureus, and Helicobacter pylori, as well as medically significant viruses.
Conclusion
Despite its considerable advantages, such as cost-effectiveness, simplicity, and the elimination of the need for specialized thermal equipment, LAMP still faces challenges. These include the complexities involved in designing multiple primers for effective gene target amplification and the limited availability of commercial kits based on the technique. Nevertheless, by addressing these drawbacks and refining the method further, LAMP has the potential to serve as a significant alternative to PCR in various laboratory settings and for rapid diagnostics in less-equipped environments. Overall, developments in molecular detection methods like LAMP signify a crucial step toward more accessible and efficient medical microbiology practices.
FAQ section (important questions/answers):
What are the main advantages of Loop Mediated Isothermal Amplification (LAMP)?
LAMP offers several advantages: it operates under isothermal conditions, requires fewer resources than PCR, and allows for visual detection of products without electrophoresis. It is also highly specific, capable of amplifying small amounts of DNA efficiently, making it accessible and user-friendly.
How does LAMP differ from traditional PCR methods?
Unlike PCR, which requires thermal cycling for DNA amplification, LAMP functions at a constant temperature. It uses multiple primers and can often yield results faster and without sophisticated equipment, making it suitable for use in less equipped laboratories.
What limitations does LAMP face compared to PCR?
LAMP's limitations include the complexity of designing multiple primers, potential difficulty in interpreting results due to diverse product sizes, and a lack of commercially available kits. These factors contribute to its lower familiarity among researchers compared to PCR.
In what applications is LAMP used for diagnostics?
LAMP is utilized for diagnosing various infectious diseases, identifying pathogens such as Mycobacterium tuberculosis, and detecting viruses. Its cost-effectiveness and simplicity make it an attractive method for laboratories aiming for rapid and reliable results.
Glossary definitions and references:
Scientific and Ayurvedic Glossary list for “LAMP Method as One of the Best Candidates for Replacing with PCR Method”. This list explains important keywords that occur in this article and links it to the glossary for a better understanding of that concept in the context of Ayurveda and other topics.
1) Lamp:
Lamp refers to Loop-Mediated Isothermal Amplification, a molecular technique for DNA amplification that circumvents the need for a thermocycler. This method is beneficial in various laboratory settings, particularly for detecting infectious agents due to its sensitivity, specificity, and ability to be executed at a constant temperature, making it more accessible than PCR.
2) Food:
Food relates to the application of Loop-Mediated Isothermal Amplification in the context of food safety. This method can be employed to detect pathogens such as Salmonella in food products, ensuring food safety and public health by identifying potential contamination quickly and accurately without requiring complex laboratory setups.
3) Aureus:
Aureus, specifically Staphylococcus aureus, is a significant pathogenic bacterium often involved in various infections. The LAMP method can rapidly detect methicillin-resistant Staphylococcus aureus (MRSA), highlighting its importance in clinical microbiology and infection control due to the bacterium's ability to cause severe health complications in humans.
4) Water:
Water signifies a critical sample matrix for microbiological testing, including the detection of pathogens using the LAMP technique. Detecting bacterial contamination in water sources is essential for public health, ensuring that drinking water is safe and free from potentially harmful microorganisms.
5) Surface:
Surface refers to the potential source of infection in studies involving LAMP. The term highlights the importance of environmental surfaces, such as skin or surfaces in healthcare settings, in the transmission of pathogens, emphasizing the relevance of efficient detection methods like LAMP in clinical diagnostics and public health.
6) Chaga:
Chaga may relate to Trypanosoma cruzi, the causative agent of Chagas disease. The LAMP method's applicability in diagnosing this disease signifies its importance in identifying parasitic infections quickly and effectively, aiding in timely treatment and intervention to prevent severe health outcomes associated with Chagas disease.
7) Kumar:
Kumar is a reference to one of the authors in the context of scientific literature on the detection of pathogens using molecular methods. Author contributions such as that of Kumar indicate the collaborative nature of research, advancing the understanding of diseases and the development of improved diagnostic techniques like PCR and LAMP.
8) Fever:
Fever in this context typically relates to typhoid fever, an infectious disease caused by Salmonella typhi. The detection of such pathogens is crucial in a clinical setting, and methods like PCR and LAMP are employed to diagnose infections accurately, allowing for prompt treatment and containment.
9) Fish:
Fish is related to the application of the LAMP technique in detecting infectious agents in aquatic environments. Specifically, this highlights the importance of molecular diagnostics in aquaculture for identifying diseases affecting fish populations, which can have significant economic implications and impact food safety.
10) Tuberculosis:
Tuberculosis, caused by Mycobacterium tuberculosis, is a contagious disease highlighting the importance of rapid diagnosis. The use of LAMP in its detection allows for prompt identification of the infection, essential for controlling outbreaks and ensuring adequate treatment to improve patient outcomes and public health.
11) Disease:
Disease refers to a broad category encompassing various health conditions, including infectious diseases where diagnostic methods like LAMP can be applied. Effective detection of pathogens is crucial in managing diseases, supporting research efforts in finding treatments, and developing preventive measures in healthcare.
12) Species:
Species refers to the classification of bacteria and other microorganisms targeted by diagnostic methods like LAMP. Identifying specific species is vital in epidemiological studies, understanding pathogen behavior, and ensuring appropriate treatment and management strategies in healthcare settings.
13) Medium:
Medium often pertains to the substance in which organisms grow or the matrix used in molecular diagnostics. In the context of LAMP, the reaction medium can define how effectively the amplification occurs, influencing the detection of pathogens and the overall efficiency of diagnostic tests.
14) Hara:
Hara is a reference to an author contributing to the scientific understanding of molecular diagnostics. Author names in research highlight the collaborative nature of scientific progress, reflecting the collective effort in developing techniques like LAMP that enhance disease detection and improve public health outcomes.
15) Post:
Post in this context may refer to the post-diagnosis stage, where the results from diagnostic methods like LAMP inform treatment decisions. Efficient post-diagnosis processes are crucial for public health management, facilitating timely interventions to control the spread of infectious diseases.
Other Health Sciences Concepts:
Discover the significance of concepts within the article: ‘LAMP Method as One of the Best Candidates for Replacing with PCR Method’. Further sources in the context of Health Sciences might help you critically compare this page with similair documents:
Infectious disease, Polymerase chain reaction, Turbidity, Toxic material, Cost-effective method, Pathogenic microorganism, Reaction mixture, PCR method, LAMP technique, Ethidium bromide, Commercial kit, PCR product, Molecular method, DNA structure, LAMP method, Optical microscope, Medical microbiology, Loop-mediated isothermal amplification, Detection method, Magnesium pyrophosphate, Target sequence, Simpler method, Oligonucleotide probes, Internal primers, External primers, Gene sequence, Reverse transcription loop-Mediated isothermal amplification.