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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®) 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.
PMID
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Authors

Mayor MeshTerms
Keywords
Journal Title plos one
Publication Year Start




PMID- 29261761
OWN - NLM
STAT- MEDLINE
DCOM- 20180108
LR  - 20180108
IS  - 1932-6203 (Electronic)
IS  - 1932-6203 (Linking)
VI  - 12
IP  - 12
DP  - 2017
TI  - Early detection of Mycobacterium avium subsp. paratuberculosis infection in
      cattle with multiplex-bead based immunoassays.
PG  - e0189783
LID - 10.1371/journal.pone.0189783 [doi]
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(R)) 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&lt;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.
FAU - Li, Lingling
AU  - Li L
AD  - Department of Animal Science, The Pennsylvania State University, University Park,
      PA, United States of America.
AD  - Huck Institutes of Life Sciences, The Pennsylvania State University, University
      Park, PA, United States of America.
FAU - Wagner, Bettina
AU  - Wagner B
AD  - Department of Population Medicine and Diagnostic Sciences, Cornell University,
      Ithaca, NY, United States of America.
FAU - Freer, Heather
AU  - Freer H
AD  - Department of Population Medicine and Diagnostic Sciences, Cornell University,
      Ithaca, NY, United States of America.
FAU - Schilling, Megan
AU  - Schilling M
AD  - Department of Animal Science, The Pennsylvania State University, University Park,
      PA, United States of America.
AD  - Huck Institutes of Life Sciences, The Pennsylvania State University, University
      Park, PA, United States of America.
FAU - Bannantine, John P
AU  - Bannantine JP
AD  - National Animal Disease Center USDA-ARS, Ames, IA, United States of America.
FAU - Campo, Joseph J
AU  - Campo JJ
AD  - Antigen Discovery, Inc., Irvine, CA, United States of America.
FAU - Katani, Robab
AU  - Katani R
AD  - Department of Animal Science, The Pennsylvania State University, University Park,
      PA, United States of America.
AD  - Huck Institutes of Life Sciences, The Pennsylvania State University, University
      Park, PA, United States of America.
FAU - Grohn, Yrjo T
AU  - Grohn YT
AD  - Department of Population Medicine and Diagnostic Sciences, Cornell University,
      Ithaca, NY, United States of America.
FAU - Radzio-Basu, Jessica
AU  - Radzio-Basu J
AD  - Huck Institutes of Life Sciences, The Pennsylvania State University, University
      Park, PA, United States of America.
FAU - Kapur, Vivek
AU  - Kapur V
AUID- ORCID: http://orcid.org/0000-0002-9648-0138
AD  - Department of Animal Science, The Pennsylvania State University, University Park,
      PA, United States of America.
AD  - Huck Institutes of Life Sciences, The Pennsylvania State University, University
      Park, PA, United States of America.
LA  - eng
PT  - Journal Article
DEP - 20171219
PL  - United States
TA  - PLoS One
JT  - PloS one
JID - 101285081
RN  - 0 (Antibodies, Bacterial)
RN  - 0 (Antigens, Bacterial)
RN  - 0 (Recombinant Proteins)
SB  - IM
MH  - Animals
MH  - Antibodies, Bacterial/blood/immunology
MH  - Antigens, Bacterial/immunology
MH  - Cattle
MH  - Cattle Diseases/blood/immunology/*microbiology
MH  - Fluorescence
MH  - Immunoassay/*methods
MH  - Milk/microbiology
MH  - Mycobacterium avium subsp. paratuberculosis/immunology/*isolation &amp; purification
MH  - Paratuberculosis/blood/immunology/*microbiology
MH  - ROC Curve
MH  - Recombinant Proteins/metabolism
MH  - Sensitivity and Specificity
MH  - Serum/microbiology
PMC - PMC5736219
EDAT- 2017/12/21 06:00
MHDA- 2018/01/09 06:00
CRDT- 2017/12/21 06:00
PHST- 2017/10/13 00:00 [received]
PHST- 2017/12/01 00:00 [accepted]
PHST- 2017/12/21 06:00 [entrez]
PHST- 2017/12/21 06:00 [pubmed]
PHST- 2018/01/09 06:00 [medline]
AID - 10.1371/journal.pone.0189783 [doi]
AID - PONE-D-17-36868 [pii]
PST - epublish
SO  - PLoS One. 2017 Dec 19;12(12):e0189783. doi: 10.1371/journal.pone.0189783.
      eCollection 2017.