BROUKHANSKI G, JAMIESON FB, WOBESER W, HOEPPNER V; Interscience Conference on Antimicrobial Agents and Chemotherapy.

Abstr Intersci Conf Antimicrob Agents Chemother Intersci Conf Antimicrob Agents Chemother. 1999 Sep 26-29; 39: 275 (abstract no. 2291).

Central Publ. Health Lab., Etobicoke, ON, CANADA

Molecular typing of M. tuberculosis (Mtb) has provided important epidemiological information for the investigation of transmission of tuberculosis. Currently, most typing is performed using the internationally standardized restriction fragment length polymorphism (RFLP) protocol, based on the insertion sequence, IS6110. We had previously examined isolates from a remote northern Canadian community (town 1), which demonstrated little or no heterogeneity between isolates using IS6110 typing. Epidemiological analysis of the clinical data did not reveal links between many of the cases with identical Mtb isolate patterns. We therefore evaluated other methods for molecular typing to determine if we could demonstrate differences or confirm the homogeneity of this group of Mtb isolates. Ten isolates from town 1 and two control isolates (Mt 14323 and Mtb H37Rv) were subjected to the following methods: spoligotyping, ERIC-PCR, BOX-PCR, and analysis of gel patterns obtained from enzymatic digestion of isolates using PvuII, Eco91I, BclI, XbaI, Bst1107I, AvrII, HinfI. Spoligotyping did not identify the minor variations seen with IS6110 RFLP. Both PCR methods, while demonstrating differences, were not reproducible. We have previously demonstrated the usefulness of PvuII for typing of Mtb strains that have significantly different IS6110 RFLP patterns, but found that PvuII cannot differentiate closely related strains. Eco91I was able to show differences between isolates that were closely related, and confirmed the homogeneity between other isolates previously demonstrated by IS6110 RFLP. Molecular typing using the enzyme Eco91I is a simple, faster and more economical alternative to IS6110 RFLP, and the resolving power of Eco91I does not depend on the number of IS6110 copies present. The patterns generated can be compared using computer assisted analysis progra ms.

Publication Types:

• Meeting Abstracts

Keywords:

• Canada
• DNA Fingerprinting
• DNA Transposable Elements
• Mycobacterium tuberculosis
• Polymerase Chain Reaction
• Polymorphism, Restriction Fragment Length
• Tuberculosis
• genetics
• instrumentation
• methods

Other ID:

• GWAIDS0007833

UI: 102245330

From Meeting Abstracts