|Title||Detection of pathogen-specific antibodies by loop-mediated isothermal amplification.|
|Publication Type||Journal Article|
|Year of Publication||2015|
|Authors||Burbulis, I. E., K. Yamaguchi, O. V. Nikolskaia, S. T. Prigge, S. Magez, S. Bisser, M. E. Reller, and D. J. Grab|
|Journal||Clin Vaccine Immunol|
|Date Published||2015 Apr|
|Keywords||Animals, Antibodies, Protozoan, Humans, Immunoglobulin G, Mice, Molecular Diagnostic Techniques, Nucleic Acid Amplification Techniques|
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.
|Alternate Journal||Clin. Vaccine Immunol.|
|PubMed Central ID||PMC4375353|
|Grant List||R01 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
Detection of pathogen-specific antibodies by loop-mediated isothermal amplification.