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Publications

When the Tuberculin Skin Test Fails: One Institution’s Experience with the Identification of TB in Research Cynomolgus Macaques.

Jay A, Marko S, Luke K, Ramos‐Rivera E, Culbreth M, Moreau A,Lugo‐Roman L, Bentzel D.  In: Association Of Primate Veterinarians 46th Annual Workshop, 2018.

While mandatory screening by administration of intradermal tuberculin skin test (TST) in captive nonhuman primate colonies has resulted in significant reduction of Mycobacterium tuberculosis (M. tb) incidence, animals imported from countries with high rates of human infection still present a significant risk to research institutes and open colonies utilizing them. At a research institute in Maryland, U.S., cynomolgus macaques were identified with M. tb. Upon discovery of the first macaque with M. tb, room and building level quarantines, changes in personal protective equipment, consideration and employment of various diagnostic screening methods, retrospective postmortem lesion analysis, and enhanced employee tuberculosis (TB) screening were conducted. A test-and-cull strategy was established for cynomolgus macaques housed in the facility utilizing standard TSTs paired with serology conducted with a commercially available simian TB test. Antemortem serology yielded 6/120 animals positive for M. tb complex (M. tb, M. bovis, M. kansasii). All macaques that were euthanized had full necropsies, histopathological evaluation, PCR testing of tissue. Of the 6 that tested positive, 1 had significant gross lesions that were positive on histopathological evaluation, PCR and culture positive  M. tb. Post-mortem serology using stored samples revealed an additional 1/10 animals positive with M. tb complex. A retrospective survey of histopathological evaluations of animals necropsied within the last 2 y revealed an additional 6 animals with granulomatous lesions and when tested for M. tb with immunofluorescent staining were positive. TS failed to identify any positive animals in the colony at any time. Improved Tb surveillance, beyond TST monitoring, should be considered for high risk groups of nonhuman primates. Incorporating serology as a component of a TB screening program for cynomolgus macaques may prevent the loss of animals, disruption of research, occupational health risk to personnel, and overall economic burden related to outbreak and disease control.

Specific pathogen free macaque colonies: a review of principles and recent advances for viral testing and colony management

JAL Yee, TH Vanderford, ES Didier… – Journal of medical …, 2016 – Wiley Online Library
Page 1. REVIEW ARTICLE Specific pathogen free macaque colonies: a review of
principles and recent advances for viral testing and colony management JoAnn L.
Yee1, Thomas H. Vanderford2, Elizabeth S. Didier3, Stanton

Novel Assay for Detecting Mycobacterium tuberculosis in Nonhuman Primates

K Luke*, JA Yee, K Andrews, D Kaushal, B Patterson Intuitive Biosciences, Madison, WI
Journal of the American Association for Laboratory Animal Science, 2013
Nonhuman primates (NHPs) used for research purposes are regularly screened for M. tuberculosis infection, both as part of transport-associated quarantine and for routine colony health maintenance programs. The current recommended screening method relies on the tuberculin skin test (TST) to identify animals with acute or latent M. tuberculosis infection; however, this method is labor intensive, takes 3 d to complete, and has known problems with false positive reactors and anergic response by infected animals. More effective testing with higher sensitivity is needed, while remaining economic in material cost and reducing labor required to perform testing on large numbers of animals. This work describes a more accurate antibody detection test for TB surveillance in NHPs by identifying antigens and using the multiplex format of the peptide microarray for simple high throughput antibody detection assay. A large set of immunoreactive antigens was first identified using peptide arrays derived from open reading frames of the M. tuberculosis genome, screening serum samples from TST test positive and negative rhesus macaques. The top immunoreactive peptides were chosen for further analysis based on ability to distinguish positive and negative samples by various analysis methods. Next, a large set of positive (both experimental and naturally occurring infections) and negative (by routine TST) serum samples from rhesus and cynomolgus macaques were run on the peptide arrays to determine sensitivity and specificity. Results from further screening at several NHP colonies with historically negative TST are reported here. Overall, this work identifies novel immunoreactive peptide antigens and an antibody detection assay for M. tuberculosis that provide high sensitivity and specificity for colony surveillance programs.

Epitope analysis of Ara h 2 and Ara h 6: characteristic patterns of IgE‐binding fingerprints among individuals with similar clinical histories

K Otsu, R Guo, SC Dreskin – Clinical & Experimental Allergy, 2015 – Wiley Online Library
Microarray printing. Triplicate spots were printed on APiX Protein Microarray Slides coated with
a proprietary optically clear nitrocellulose (Intuitive Biosciences, Madison, WI, USA). All slides
were stored at 4°C and used within 6 months from the date of manufacture.

Identification and spontaneous immune targeting of an endogenous retrovirus K envelope protein in the Indian rhesus macaque model of human disease

HL Wu, EJ Léon, LT Wallace… – …, 2016 – retrovirology.biomedcentral.com
ELISA was conducted in duplicate, using absorbance at 450 nm to determine binding
antibodies. Serological tests for antibodies. Intuitive Biosciences performed serology
assays. Individual serum samples were screened for

ChIP-less analysis of chromatin states

Z Su, MD Boersma, JH Lee… – Epigenetics & …, 2014 – epigeneticsandchromatin. …
Each solution was mixed in separate wells of 384-well polypropylene plates and
spotted in triplicate onto 75 mm × 25 mm nitrocellulose sides (Intuitive Biosciences)
with a Gene Machines OmniGrid Arrayer (Genomic Solutions).