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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.
Am J Gastroenterol 2005; 100(7):1529-36AJ

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.

Authors+Show Affiliations

Dipartimento di Scienze Biomediche, Sezione di Microbiologia Sperimentale e Clinica, Università degli studi di Sassari, Sassari, Italy.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

15984976

Citation

Sechi, Leonardo A., et al. "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, no. 7, 2005, pp. 1529-36.
Sechi LA, Scanu AM, Molicotti P, et al. Detection and Isolation of Mycobacterium avium subspecies paratuberculosis from intestinal mucosal biopsies of patients with and without Crohn's disease in Sardinia. Am J Gastroenterol. 2005;100(7):1529-36.
Sechi, L. A., Scanu, A. M., Molicotti, P., Cannas, S., Mura, M., Dettori, G., ... Zanetti, S. (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, 100(7), pp. 1529-36.
Sechi LA, et al. Detection and Isolation of Mycobacterium Avium Subspecies Paratuberculosis From Intestinal Mucosal Biopsies of Patients With and Without Crohn's Disease in Sardinia. Am J Gastroenterol. 2005;100(7):1529-36. PubMed PMID: 15984976.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Detection and Isolation of Mycobacterium avium subspecies paratuberculosis from intestinal mucosal biopsies of patients with and without Crohn's disease in Sardinia. AU - Sechi,Leonardo A, AU - Scanu,Antonio M, AU - Molicotti,Paola, AU - Cannas,Sara, AU - Mura,Manuela, AU - Dettori,Giuseppe, AU - Fadda,Giovanni, AU - Zanetti,Stefania, PY - 2005/6/30/pubmed PY - 2005/9/1/medline PY - 2005/6/30/entrez SP - 1529 EP - 36 JF - The American journal of gastroenterology JO - Am. J. Gastroenterol. VL - 100 IS - 7 N2 - 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. SN - 0002-9270 UR - https://www.unboundmedicine.com/medline/citation/15984976/Detection_and_Isolation_of_Mycobacterium_avium_subspecies_paratuberculosis_from_intestinal_mucosal_biopsies_of_patients_with_and_without_Crohn's_disease_in_Sardinia_ DB - PRIME DP - Unbound Medicine ER -