Early detection of Mycobacterium avium subsp. paratuberculosis infection in cattle with multiplex-bead based immunoassays

Lingling Li, Bettina Wagner, Heather Freer, Megan Schilling, John P. Bannantine, Joseph J. Campo, Robab Katani, Yrjo T. Grohn, Jessica Radzio-Basu, Vivek Kapur

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Johne’s Disease (JD), caused by Mycobacterium avium subspecies paratuberculosis (MAP), results in significant economic loss to livestock production. The early detection of MAP infection in animals with extant serological assays has remained challenging due to the low sensitivity of commercially available ELISA tests, a fact that has hampered the development of effective JD control programs. Our recent protein microarray-based studies identified several promising candidate antigens that are immunogenic during different stages of MAP infection. To evaluate these antigens for use in diagnostic assays and reliably identify animals with MAP infection, a multiplex (Luminex ® ) assay was developed using color-coded flourescent beads coupled to 6 MAP recombinant proteins and applied to screen 180 serum and 90 milk samples from cows at different stages of MAP infection including negative (NL), fecal test positive/ELISA negative (F+E-), and fecal positive/ELISA positive (F+E+). The results show that while serum antibody reactivities to each of the 6 antigens were highest in F+E+ group, antibody reactivity to three of the six antigens were identified in the F+E- group, suggesting that these three antigens are expressed and provoke antibody responses during the early infection stages with MAP. Further, antibodies against all six antigens were elevated in milk samples from both the F+E- and F+E+ groups in comparison to the NL group (p<0.01). Taken together, the results of our investigation suggest that multiplex bead-based assays are able to reliably identify MAP infection, even during early stages when antibody responses in animals are undetectable with widely used commercial ELISA tests.

Original languageEnglish (US)
Article numbere0189783
JournalPloS one
Volume12
Issue number12
DOIs
StatePublished - Dec 2017

Fingerprint

Mycobacterium avium subsp. paratuberculosis
Paratuberculosis
Mycobacterium avium
paratuberculosis
immunoassays
Immunoassay
Antigens
cattle
Assays
Infection
Antibodies
infection
antigens
Animals
Enzyme-Linked Immunosorbent Assay
antibodies
enzyme-linked immunosorbent assay
Disease control
Antibody Formation
assays

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Li, Lingling ; Wagner, Bettina ; Freer, Heather ; Schilling, Megan ; Bannantine, John P. ; Campo, Joseph J. ; Katani, Robab ; Grohn, Yrjo T. ; Radzio-Basu, Jessica ; Kapur, Vivek. / Early detection of Mycobacterium avium subsp. paratuberculosis infection in cattle with multiplex-bead based immunoassays. In: PloS one. 2017 ; Vol. 12, No. 12.
@article{2dbc7cc7b7e54bd7963f4d56ea222148,
title = "Early detection of Mycobacterium avium subsp. paratuberculosis infection in cattle with multiplex-bead based immunoassays",
abstract = "Johne’s Disease (JD), caused by Mycobacterium avium subspecies paratuberculosis (MAP), results in significant economic loss to livestock production. The early detection of MAP infection in animals with extant serological assays has remained challenging due to the low sensitivity of commercially available ELISA tests, a fact that has hampered the development of effective JD control programs. Our recent protein microarray-based studies identified several promising candidate antigens that are immunogenic during different stages of MAP infection. To evaluate these antigens for use in diagnostic assays and reliably identify animals with MAP infection, a multiplex (Luminex {\circledR} ) assay was developed using color-coded flourescent beads coupled to 6 MAP recombinant proteins and applied to screen 180 serum and 90 milk samples from cows at different stages of MAP infection including negative (NL), fecal test positive/ELISA negative (F+E-), and fecal positive/ELISA positive (F+E+). The results show that while serum antibody reactivities to each of the 6 antigens were highest in F+E+ group, antibody reactivity to three of the six antigens were identified in the F+E- group, suggesting that these three antigens are expressed and provoke antibody responses during the early infection stages with MAP. Further, antibodies against all six antigens were elevated in milk samples from both the F+E- and F+E+ groups in comparison to the NL group (p<0.01). Taken together, the results of our investigation suggest that multiplex bead-based assays are able to reliably identify MAP infection, even during early stages when antibody responses in animals are undetectable with widely used commercial ELISA tests.",
author = "Lingling Li and Bettina Wagner and Heather Freer and Megan Schilling and Bannantine, {John P.} and Campo, {Joseph J.} and Robab Katani and Grohn, {Yrjo T.} and Jessica Radzio-Basu and Vivek Kapur",
year = "2017",
month = "12",
doi = "10.1371/journal.pone.0189783",
language = "English (US)",
volume = "12",
journal = "PLoS One",
issn = "1932-6203",
publisher = "Public Library of Science",
number = "12",

}

Early detection of Mycobacterium avium subsp. paratuberculosis infection in cattle with multiplex-bead based immunoassays. / Li, Lingling; Wagner, Bettina; Freer, Heather; Schilling, Megan; Bannantine, John P.; Campo, Joseph J.; Katani, Robab; Grohn, Yrjo T.; Radzio-Basu, Jessica; Kapur, Vivek.

In: PloS one, Vol. 12, No. 12, e0189783, 12.2017.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Early detection of Mycobacterium avium subsp. paratuberculosis infection in cattle with multiplex-bead based immunoassays

AU - Li, Lingling

AU - Wagner, Bettina

AU - Freer, Heather

AU - Schilling, Megan

AU - Bannantine, John P.

AU - Campo, Joseph J.

AU - Katani, Robab

AU - Grohn, Yrjo T.

AU - Radzio-Basu, Jessica

AU - Kapur, Vivek

PY - 2017/12

Y1 - 2017/12

N2 - Johne’s Disease (JD), caused by Mycobacterium avium subspecies paratuberculosis (MAP), results in significant economic loss to livestock production. The early detection of MAP infection in animals with extant serological assays has remained challenging due to the low sensitivity of commercially available ELISA tests, a fact that has hampered the development of effective JD control programs. Our recent protein microarray-based studies identified several promising candidate antigens that are immunogenic during different stages of MAP infection. To evaluate these antigens for use in diagnostic assays and reliably identify animals with MAP infection, a multiplex (Luminex ® ) assay was developed using color-coded flourescent beads coupled to 6 MAP recombinant proteins and applied to screen 180 serum and 90 milk samples from cows at different stages of MAP infection including negative (NL), fecal test positive/ELISA negative (F+E-), and fecal positive/ELISA positive (F+E+). The results show that while serum antibody reactivities to each of the 6 antigens were highest in F+E+ group, antibody reactivity to three of the six antigens were identified in the F+E- group, suggesting that these three antigens are expressed and provoke antibody responses during the early infection stages with MAP. Further, antibodies against all six antigens were elevated in milk samples from both the F+E- and F+E+ groups in comparison to the NL group (p<0.01). Taken together, the results of our investigation suggest that multiplex bead-based assays are able to reliably identify MAP infection, even during early stages when antibody responses in animals are undetectable with widely used commercial ELISA tests.

AB - Johne’s Disease (JD), caused by Mycobacterium avium subspecies paratuberculosis (MAP), results in significant economic loss to livestock production. The early detection of MAP infection in animals with extant serological assays has remained challenging due to the low sensitivity of commercially available ELISA tests, a fact that has hampered the development of effective JD control programs. Our recent protein microarray-based studies identified several promising candidate antigens that are immunogenic during different stages of MAP infection. To evaluate these antigens for use in diagnostic assays and reliably identify animals with MAP infection, a multiplex (Luminex ® ) assay was developed using color-coded flourescent beads coupled to 6 MAP recombinant proteins and applied to screen 180 serum and 90 milk samples from cows at different stages of MAP infection including negative (NL), fecal test positive/ELISA negative (F+E-), and fecal positive/ELISA positive (F+E+). The results show that while serum antibody reactivities to each of the 6 antigens were highest in F+E+ group, antibody reactivity to three of the six antigens were identified in the F+E- group, suggesting that these three antigens are expressed and provoke antibody responses during the early infection stages with MAP. Further, antibodies against all six antigens were elevated in milk samples from both the F+E- and F+E+ groups in comparison to the NL group (p<0.01). Taken together, the results of our investigation suggest that multiplex bead-based assays are able to reliably identify MAP infection, even during early stages when antibody responses in animals are undetectable with widely used commercial ELISA tests.

UR - http://www.scopus.com/inward/record.url?scp=85038961282&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85038961282&partnerID=8YFLogxK

U2 - 10.1371/journal.pone.0189783

DO - 10.1371/journal.pone.0189783

M3 - Article

C2 - 29261761

AN - SCOPUS:85038961282

VL - 12

JO - PLoS One

JF - PLoS One

SN - 1932-6203

IS - 12

M1 - e0189783

ER -