Q:

6/19/2014
IS Loop Mediated Isothermal Amplification (LAMP) a viable concept for creating a vaccine to eliminate various food allergies? Like peanut etc. and the current status in time that the treatment would be available. Which companies are currently conducting research in this area utilizing LAMP? Do you think it can work?

A:

Thank you for your inquiry.

Loop Mediated Isothermal Amplification (LAMP) is a technique used for the rapid amplification of nucleic acid. It is therefore similar in purpose to the older polymerase chain reaction (PCR). It has advantages and disadvantages when compared to a PCR, but at the present time, its uses are similar to those for PCR. Its major application is for the detection of the presence of infectious organisms. It has been used to detect a number of these including tuberculosis, sleeping sickness, and recently Strongyloides (see abstract copied below). A review of its technology and uses can be seen in the Parida M, et al., abstract copied below.

More recently, it has been employed to detect food allergens (see Zahradnik, et al., copied below). One of the companies dealing with LAMP is Lucigen. I have copied below a link to their website. Their contact information can be found there.

Finally, there is a very well done PowerPoint presentation on LAMP by Dr. Panagiotis Karanis here.

I suppose a future use of LAMP could be the production of DNA to be used in vaccines, but I am not aware of any work on this at the present time, and could find no evidence for such research on an Internet search. This of course does not mean that work on DNA vaccines that employ LAMP to produce the DNA is not being done, but if so, my opinion is that it would be in a very early stage and the process would be far too premature, at least in my opinion, to assess its utility and future. I think it is important to remember that this is not a unique technique. We have been performing rapid DNA amplication for many years. This technique, as noted, have some advantages over PCR. However, I could find nothing in the literature which would support this technique to represent a unique advance in the attempt to develop a vaccine for food allergy.

Thank you again for your inquiry and we hope this response is helpful to you.

Am J Trop Med Hyg. 2014 Feb;90(2):306-11. doi: 10.4269/ajtmh.13-0583. Epub 2013 Dec 9.
A loop-mediated isothermal amplification (LAMP) assay for Strongyloides stercoralis in stool that uses a visual detection method with SYTO-82 fluorescent dye.
Watts MR1, James G, Sultana Y, Ginn AN, Outhred AC, Kong F, Verweij JJ, Iredell JR, Chen SC, Lee R.
Author information
1Centre for Infectious Diseases and Microbiology Public Health; Pathology West - Institute of Clinical Pathology and Medical Research, Westmead Hospital, New South Wales, Australia; Marie Bashir Institute for Infectious Diseases and Biosecurity and Centre for Research Excellence in Critical Infection, University of Sydney, New South Wales, Australia; Department of Zoology, University of Dhaka, Dhaka, Bangladesh; Department of Parasitology, Leiden University Medical Center, Leiden, The Netherlands.
Abstract
An assay to detect Strongyloides stercoralis in stool specimens was developed using the loop-mediated isothermal amplification (LAMP) method. Primers were based on the 28S ribosomal subunit gene. The reaction conditions were optimized and SYTO-82 fluorescent dye was used to allow real-time and visual detection of the product. The product identity was confirmed with restriction enzyme digestion, cloning, and sequence analysis. The assay was specific when tested against DNA from bacteria, fungi and parasites, and 30 normal stool samples. Analytical sensitivity was to < 10 copies of target sequence in a plasmid and up to a 10(-2) dilution of DNA extracted from a Strongyloides ratti larva spiked into stool. Sensitivity was increased when further dilutions were made in water, indicative of reduced reaction inhibition. Twenty-seven of 28 stool samples microscopy and polymerase chain reaction positive for S. stercoralis were positive with the LAMP method. On the basis of these findings, the assay warrants further clinical validation.

Rev Med Virol. 2008 Nov-Dec;18(6):407-21. doi: 10.1002/rmv.593.
Loop mediated isothermal amplification (LAMP): a new generation of innovative gene amplification technique; perspectives in clinical diagnosis of infectious diseases.
Parida M1, Sannarangaiah S, Dash PK, Rao PV, Morita K.
Author information
1Division of Virology, Defence Research & Development Establishment, Gwalior 474002, MP, India.
Abstract
Loop mediated isothermal amplification (LAMP) is a powerful innovative gene amplification technique emerging as a simple rapid diagnostic tool for early detection and identification of microbial diseases. The whole procedure is very simple and rapid wherein the amplification can be completed in less than 1 h under isothermal conditions employing a set of six specially designed primers spanning eight distinct sequences of a target gene, by incubating all the reagents in a single tube. Gene amplification products can be detected by agarose gel electrophoresis as well as by real-time monitoring in an inexpensive turbidimeter. Gene copy number can also be quantified with the help of a standard curve generated from different concentrations of gene copy number plotted against time of positivity with the help of a real-time turbidimeter. Alternatively, gene amplification can be visualised by the naked eye either as turbidity or in the form of a colour change when SYBR Green I, a fluorescent dsDNA intercalating dye, is employed. LAMP does not require a thermal cycler and can be performed simply with a heating block and/or water bath. Considering the advantages of rapid amplification, simple operation and easy detection, LAMP has potential applications for clinical diagnosis as well as surveillance of infectious diseases in developing countries without requiring sophisticated equipment or skilled personnel.

Anal Bioanal Chem. 2014 Jun 1. [Epub ahead of print]
Detection of the food allergen celery via loop-mediated isothermal amplification technique.
Zahradnik C1, Martzy R, Mach RL, Krska R, Farnleitner AH, Brunner K.
Author information
1Institute of Chemical Engineering, IFA-Tulln, Center for Analytical Chemistry, Vienna University of Technology, Konrad Lorenz Str. 20, 3430, Tulln, Austria.
Abstract
Since 2005, celery and celery products have to be labeled according to Directive 2003/89/EC due to their allergenic potential. In order to provide a DNA-based, rapid and simple detection method suitable for high-throughput analysis, a loop-mediated isothermal amplification (LAMP) assay for the detection of celery (Apium graveolens) was developed. The assay was tested for specificity for celery since closely related species also hold food relevance. The limit of detection (LOD) for spiked food samples was found to be as low as 7.8 mg of dry celery powder per kilogram. An evaluation of different amplification and detection platforms was performed to show reliable detection independent from the instrument used for amplification (thermal cycler or heating block) and detection mechanisms (real-time fluorescence detection, agarose gel electrophoresis or nucleic acid staining). The analysis of 10 commercial food samples representing diverse and complex food matrices, and a false-negative rate of 0 % for approximately 24 target copies or 0.08 ng celery DNA for three selected food matrices show that LAMP has the potential to be used as an alternative strategy for the detection of allergenic celery. The performance of the developed LAMP assay turned out to be equal or superior to the best available PCR assay for the detection of celery in food products.

LUCIGEN

Sincerely,
Phil Lieberman, M.D.
 

AAAAI - American Academy of Allergy Asthma & Immunology