Mycobacterium avium ss paratuberculosis In Crohn’s Disease Tissue Is an Affirmative!
D.Schwartz,
C. Romero, D. Campbell, I. Shafran and S. Naser
Department
of Molecular Biology and Microbiology/ Center for Discovery of Drugs and
Diagnostics
University
of Central Florida, Orlando, Florida
The insertion sequence IS900 of Mycobacterium avium ss paratuberculosis (M para) has been used as a fingerprint-type marker for
identification of M para or M para-like microorganisms in clinical specimens from patients with
Crohn’s disease (CD), a chronic inflammatory bowel disease. However,
amplification of IS900 in these specimens does not confirm the virulent form of
the bacterium or the status of disease. Culturing M para from CD tissue has been reported but with limited success.
This, in part, is significantly related to the type of culture media and the
integrity of the specimen used. Amplification of the IS900 followed by DNA
hybridization from cultured tissue, provide accurate diagnosis of CD. In
this study, 59 tissue specimens from 59 patients (20 CD consisting of 7 resected
tissue and 13 biopsies and 39 control consisting of 1 resected tissue and 38
biopsies from Ulcerative colitis and noninflammatory bowel disease patients)
were homogenized, decontaminated and inoculated into BACTEC bottles and MGIT
media (radioactive and non-radioactive methods, respectively). Monthly aliquots
of incubated cultures were examined microscopically and by PCR/hybridization
using IS900-based primers for the presence of M
para. Acid fast staining and immunoscreeening using rabbit poly clonal anti-M para antibodies by confocal laser microscopy revealed the presence
of M para in 6 CD specimens. The
positive specimens consisted of 6 resected tissue and 1 biopsy. This was
confirmed with PCR and hybrodization of the 39 control specimens, none were
positive with any of the assays. Our data also suggest that M
para may be isolated from tissue specimens, especially those resected (if
present) within weeks in the MGIT media.
All content copyright ©1999 Dr. Ira Shafran, M.D.