Detection of pathogen-specific antibodies by loop-mediated isothermal amplification.

TitleDetection of pathogen-specific antibodies by loop-mediated isothermal amplification.
Publication TypeJournal Article
Year of Publication2015
AuthorsBurbulis, I. E., K. Yamaguchi, O. V. Nikolskaia, S. T. Prigge, S. Magez, S. Bisser, M. E. Reller, and D. J. Grab
JournalClin Vaccine Immunol
Volume22
Issue4
Pagination374-80
Date Published2015 Apr
ISSN1556-679X
KeywordsAnimals, Antibodies, Protozoan, Humans, Immunoglobulin G, Mice, Molecular Diagnostic Techniques, Nucleic Acid Amplification Techniques
Abstract

Loop-mediated isothermal amplification (LAMP) is a method for enzymatically replicating DNA that has great utility for clinical diagnosis at the point of care (POC), given its high sensitivity, specificity, speed, and technical requirements (isothermal conditions). Here, we adapted LAMP for measuring protein analytes by creating a protein-DNA fusion (referred to here as a "LAMPole") that attaches oligonucleotides (LAMP templates) to IgG antibodies. This fusion consists of a DNA element covalently bonded to an IgG-binding polypeptide (protein L/G domain). In our platform, LAMP is expected to provide the most suitable means for amplifying LAMPoles for clinical diagnosis at the POC, while quantitative PCR is more suitable for laboratory-based quantification of antigen-specific IgG abundance. As proof of concept, we measured serological responses to a protozoan parasite by quantifying changes in solution turbidity in real time. We observed a >6-log fold difference in signal between sera from vaccinated versus control mice and in a clinical patient sample versus a control. We assert that LAMPoles will be useful for increasing the sensitivity of measuring proteins, whether it be in a clinical laboratory or in a field setting, thereby improving acute diagnosis of a variety of infections.

DOI10.1128/CVI.00811-14
Alternate JournalClin. Vaccine Immunol.
PubMed ID25651920
PubMed Central IDPMC4375353
Grant ListR01 AI082695 / AI / NIAID NIH HHS / United States
R21 AI079282 / AI / NIAID NIH HHS / United States
1R21AI079282 / AI / NIAID NIH HHS / United States
5R01AI082695 / AI / NIAID NIH HHS / United States
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