A Burkholderia pseudomallei protein microarray reveals serodiagnostic and cross-reactive antigens.
Understanding the way in which the immune system responds to infection is central to the development of vaccines and many diagnostics. To provide insight into this area, we fabricated a protein microarray containing 1,205 Burkholderia pseudomallei proteins, probed it with 88 melioidosis patient sera, and identified 170 reactive antigens. This subset of antigens was printed on a smaller array and probed with a collection of 747 individual sera derived from 10 patient groups including melioidosis patients from Northeast Thailand and Singapore, patients with different infections, healthy individuals from the USA, and from endemic and nonendemic regions of Thailand. We identified 49 antigens that are significantly more reactive in melioidosis patients than healthy people and patients with other types of bacterial infections. We also identified 59 cross-reactive antigens that are equally reactive among all groups, including healthy controls from the USA. Using these results we were able to devise a test that can classify melioidosis positive and negative individuals with sensitivity and specificity of 95% and 83%, respectively, a significant improvement over currently available diagnostic assays. Half of the reactive antigens contained a predicted signal peptide sequence and were classified as outer membrane, surface structures or secreted molecules, and an additional 20% were associated with pathogenicity, adaptation or chaperones. These results show that microarrays allow a more comprehensive analysis of the immune response on an antigen-specific, patient-specific, and population-specific basis, can identify serodiagnostic antigens, and contribute to a more detailed understanding of immunogenicity to this pathogen.
Felgner
Philip L
PL
Department of Medicine, Division of Infectious Diseases, University of California, Irvine, CA 92697, USA. pfelgner@uci.edu
Kayala
Matthew A
MA
Vigil
Adam
A
Burk
Chad
C
Nakajima-Sasaki
Rie
R
Pablo
Jozelyn
J
Molina
Douglas M
DM
Hirst
Siddiqua
S
Chew
Janet S W
JS
Wang
Dongling
D
Tan
Gladys
G
Duffield
Melanie
M
Yang
Ron
R
Neel
Julien
J
Chantratita
Narisara
N
Bancroft
Greg
G
Lertmemongkolchai
Ganjana
G
Davies
D Huw
DH
Baldi
Pierre
P
Peacock
Sharon
S
Titball
Richard W
RW
D000942:Antigens, Bacterial / Q000145:classification / Q000276:immunology*; D001426:Bacterial Proteins / Q000276:immunology*; D016957:Burkholderia pseudomallei / Q000276:immunology*; D016022:Case-Control Studies; D003429:Cross Reactions / Q000276:immunology; D018604:Epitope Mapping; D006801:Humans; D008554:Melioidosis / Q000175:diagnosis / Q000276:immunology; D040081:Protein Array Analysis; D012698:Serologic Tests; D012846:Singapore; D013785:Thailand; D014481:United States
D000942:Antigens, Bacterial; D001426:Bacterial Proteins