Skip to main content

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

Publication ,  Journal Article
Burbulis, IE; Yamaguchi, K; Nikolskaia, OV; Prigge, ST; Magez, S; Bisser, S; Reller, ME; Grab, DJ
Published in: Clin Vaccine Immunol
April 2015

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.

Duke Scholars

Altmetric Attention Stats
Dimensions Citation Stats

Published In

Clin Vaccine Immunol

DOI

EISSN

1556-679X

Publication Date

April 2015

Volume

22

Issue

4

Start / End Page

374 / 380

Location

United States

Related Subject Headings

  • Nucleic Acid Amplification Techniques
  • Molecular Diagnostic Techniques
  • Microbiology
  • Mice
  • Immunology
  • Immunoglobulin G
  • Humans
  • Antibodies, Protozoan
  • Animals
  • 3204 Immunology
 

Citation

APA
Chicago
ICMJE
MLA
NLM
Burbulis, I. E., Yamaguchi, K., Nikolskaia, O. V., Prigge, S. T., Magez, S., Bisser, S., … Grab, D. J. (2015). Detection of pathogen-specific antibodies by loop-mediated isothermal amplification. Clin Vaccine Immunol, 22(4), 374–380. https://doi.org/10.1128/CVI.00811-14
Burbulis, Ian E., Kumiko Yamaguchi, Olga V. Nikolskaia, Sean T. Prigge, Stefan Magez, Sylvie Bisser, Megan E. Reller, and Dennis J. Grab. “Detection of pathogen-specific antibodies by loop-mediated isothermal amplification.Clin Vaccine Immunol 22, no. 4 (April 2015): 374–80. https://doi.org/10.1128/CVI.00811-14.
Burbulis IE, Yamaguchi K, Nikolskaia OV, Prigge ST, Magez S, Bisser S, et al. Detection of pathogen-specific antibodies by loop-mediated isothermal amplification. Clin Vaccine Immunol. 2015 Apr;22(4):374–80.
Burbulis, Ian E., et al. “Detection of pathogen-specific antibodies by loop-mediated isothermal amplification.Clin Vaccine Immunol, vol. 22, no. 4, Apr. 2015, pp. 374–80. Pubmed, doi:10.1128/CVI.00811-14.
Burbulis IE, Yamaguchi K, Nikolskaia OV, Prigge ST, Magez S, Bisser S, Reller ME, Grab DJ. Detection of pathogen-specific antibodies by loop-mediated isothermal amplification. Clin Vaccine Immunol. 2015 Apr;22(4):374–380.

Published In

Clin Vaccine Immunol

DOI

EISSN

1556-679X

Publication Date

April 2015

Volume

22

Issue

4

Start / End Page

374 / 380

Location

United States

Related Subject Headings

  • Nucleic Acid Amplification Techniques
  • Molecular Diagnostic Techniques
  • Microbiology
  • Mice
  • Immunology
  • Immunoglobulin G
  • Humans
  • Antibodies, Protozoan
  • Animals
  • 3204 Immunology