PCR-restriction enzyme analysis of a bone marrow isolate from a human immunodeficiency virus-positive patient discloses polyclonal infection with two Mycobacterium avium strains

Abstract

Polyclonal infection by Mycobacterium avium was detected by hsp65 PCR-restriction enzyme analysis (PRA) in a bone marrow isolate from an AIDS patient. Two M. avium strains, differing in colony morphology, PRA HaeIII digestion pattern, insertion element (IS) 1245 amplification, and restriction fragment length polymorphism fingerprints with IS1245 and IS1311 probes, were isolated.

FIG. 1

Alignment of hsp65 sequences (total length, 380 positions) from OP colony-forming bacilli, TL colony-forming bacilli, M. avium ATCC 25291, and M. intracellulare ATCC 13950. Nucleotide substitutions in relation to the M. avium sequence are underlined. HaeIII sites are boxed, and asterisks indicate sites with point mutations.

FIG. 2

RFLP with DNA isolated from five OP and five TL colony-forming bacilli. The same membrane was hybridized with peroxidase-labeled IS1245 and IS1311 probes and developed with the ECL labeling and detection kit (Amersham, Pharmacia Biotech, Inc., Piscataway, N.J.). Normalization was performed with GelCompar II software (Applied Maths, Kortrijk, Belgium).