Crohn's disease: failure of a proprietary fluorescent in situ hybridization assay to detect M. avium subspecies paratuberculosis in archived frozen intestine from patients with Crohn's disease.

Autor: Greenstein RJ; Department of Surgery, James J. Peters Veterans Affairs Medical Center Bronx, New York, USA. BGAxis@aol.com.; Laboratory of Molecular Surgical Research, James J. Peters Veterans Affairs Medical Center Bronx, New York, USA. BGAxis@aol.com., Su L; Laboratory of Molecular Surgical Research, James J. Peters Veterans Affairs Medical Center Bronx, New York, USA., Fam PS; Mental Illness Research Education and Clinical Center (MIRECC), James J. Peters, Veterans Affairs Medical Center Bronx, New York, USA., Gurland B; Colorectal Surgery, Stanford University School of Medicine, Stanford CA, USA., Endres P; Department of Pathology, Icahn School of Medicine at Mount Sinai New York, New York, USA., Brown ST; Infectious Disease Section, James J. Peters Veterans Affairs Medical Center Bronx, New York, USA.; Department of Medicine, Icahn School of Medicine at Mt. Sinai. New York, New York, USA.
Jazyk: angličtina
Zdroj: BMC research notes [BMC Res Notes] 2020 Feb 24; Vol. 13 (1), pp. 96. Date of Electronic Publication: 2020 Feb 24.
DOI: 10.1186/s13104-020-04947-0
Abstrakt: Objectives: Although controversial, there is increasing concern that Crohn's disease may be a zoonotic infectious disease consequent to a mycobacterial infection. The most plausible candidate is M. avium subspecies paratuberculosis (MAP) that is unequivocally responsible for Johne's disease in ruminants. The purpose of this study was to evaluate a proprietary (Affymetrix™ RNA view ® ) fluorescent in situ hybridization (FISH) assay for MAP RNA. Non-identifiable intestine from patients with documented Crohn's disease was assayed according to the manufacturer's instructions and with suggested modifications. Probes were custom designed for MAP and human β-actin (as the eukaryotic housekeeping gene) from published genomes.
Results: Repetitively, false positive signal was observed in our "No-Probe" negative control. Attempts were made to correct this according to the manufacturer's suggestions (by modifying wash solutions, using recommended hydrochloric acid titration and different fluorescent filters). None prevented false positive signal in the "No-Probe" control. It is concluded that when performed according to manufactures instruction and with multiple variations on the manufactures recommended suggestions to correct for false positive signal, that the Affymetrix™ RNA view ® cannot be used to detect MAP in pre-frozen resected intestine of humans with Crohn's disease.
Databáze: MEDLINE
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