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

Ian E. Burbulis, Kumiko Yamaguchi, Olga V. Nikolskaia, Sean T. Prigge, Stefan Magez, Sylvie Bisser, Megan E. Reller, Dennis J. Grab

Research output: Contribution to journalArticle

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 laboratorybased 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.

Original languageEnglish (US)
Pages (from-to)374-380
Number of pages7
JournalClinical and Vaccine Immunology
Volume22
Issue number4
DOIs
StatePublished - Apr 1 2015

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology
  • Clinical Biochemistry
  • Microbiology (medical)

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  • Cite this

    Burbulis, I. E., Yamaguchi, K., Nikolskaia, O. V., Prigge, S. T., Magez, S., Bisser, S., Reller, M. E., & Grab, D. J. (2015). Detection of pathogen-specific antibodies by loop-mediated isothermal amplification. Clinical and Vaccine Immunology, 22(4), 374-380. https://doi.org/10.1128/CVI.00811-14