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Detection and isolation of Mycobacterium avium subspecies paratuberculosis from intestinal mucosal biopsies of patients with and without Crohn's disease in Sardinia

Sechi, Leonardo Antonio and Scanu, Antonio Mario and Molicotti, Paola and Cannas, Sara and Mura, Manuela and Dettori, Giuseppe Lorenzo and Fadda, Giovanni and Zanetti, Stefania Anna Lucia (2005) Detection and isolation of Mycobacterium avium subspecies paratuberculosis from intestinal mucosal biopsies of patients with and without Crohn's disease in Sardinia. The American Journal of Gastroenterology, Vol. 100 (7), p. 1529-1536. eISSN 1572-0241. Article.

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DOI: 10.1111/j.1572-0241.2005.41415.x

Abstract

OBJECTIVES: Sardinia is an island community of 1.6 million people. There are also about 3.5 million sheep and one hundred thousand cattle in which Johne's disease and Mycobacterium avium subspecies paratuberculosis infection are endemic. The present study was designed to determine what proportion of people in Sardinia attending for ileocolonoscopy with or without Crohn's disease were infected with this pathogen.
METHODS: Mycobacterium avium subspecies paratuberculosis was detected by IS900 PCR on DNA extracts of fresh intestinal mucosal biopsies as well as by isolation in culture using supplemented MGIT media followed by PCR with amplicon sequencing.
RESULTS: Twenty five patients (83.3%) with Crohn's disease and 3 control patients (10.3%) were IS900 PCR positive (p = 0.000001; Odds ratio 43.3). Mycobacterium avium subspecies paratuberculosis grew in cultures from 19 Crohn's patients (63.3%) and from 3 control patients (10.3%) (p = 0.00001; Odds ratio 14.9). All patients positive by culture had previously been positive by PCR. Mycobacterium avium subspecies paratuberculosis first appeared in the liquid cultures in a Ziehl Neelsen (ZN) staining negative form and partially reverted through a rhodamine-auramine positive staining form to the classical ZN positive form. This resulted in a stable mixed culture of all 3 forms illustrating the phenotypic versatility of these complex chronic enteric pathogens.
CONCLUSIONS: Mycobacterium avium subspecies paratuberculosis was detected in the majority of Sardinian Crohn's disease patients. The finding of the organism colonizing a proportion of people without Crohn's disease is consistent with what occurs in other conditions caused by a primary bacterial pathogen in susceptible hosts.

Item Type:Article
ID Code:3858
Status:Published
Refereed:Yes
Uncontrolled Keywords:Mycobacterium avium subspecies paratuberculosis, Johne's disease, Crohn's disease, Sardinia
Subjects:Area 06 - Scienze mediche > MED/07 Microbiologia e microbiologia clinica
Area 06 - Scienze mediche > MED/18 Chirurgia generale
Divisions:001 Università di Sassari > 03 Istituti > Clinica chirurgica generale e terapia chirurgica
001 Università di Sassari > 01 Dipartimenti > Scienze biomediche
002 Altri enti e centri di ricerca del Nord Sardegna > ISOGEM-International Society for Genomic and Evolutionary Microbiology, Sassari
Publisher:Nature Publishing Group
eISSN:1572-0241
Copyright Holders:© 2005 The American College of Gastroenterology
Deposited On:11 May 2010 13:07

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