Restriction Fragment Length Polymorphism Analysis of Epidemiologically Related Mycobacterium tuberculosis Isolates

Abstract

Restriction fragment length polymorphism (RFLP) analysis of a large number of Japanese isolates of Mycobacterium tuberculosis, containing isolates from small outbreaks of M. tuberculosis infection, and clinical isolates of M. bovis BCG, was carried out using a DNA probe derived from the insertion sequence IS986. Clinical isolates of M. tuberculosis had a high degree of RFLP. The occurrences of the IS element varied from 1 to 19, the majority of isolates having 8 to 15 copies. Very similar fingerprints, however, were seen among strains isolated in the Kanto district. In particular, 3 strains were of the same pattern with or without an additional band. Similarity of the banding patterns of strains islated in the same district was observed in other areas. Six groups of strains, each group arising from a suspected common source of infection, were analyzed. Of these, 5 showed identical fingerprints within each group, but one showed different fingerprints. RFLP patterns of three strains isolated from individuals with lymphadenitis developed about two months after BCG vaccination, and one strain isolated from a bladder cancer patient with BCG instillation therapy were identical to those of BCG-Tokyo which had been used for the vaccination and therapy. These results confirm that RFLP analysis using IS986 is a suitable tool for epidemiology of tuberculosis.

Simple and Rapid Identification of the Mycobacterium tuberculosis Complex by Immunochromatographic Assay Using Anti-MPB64 Monoclonal Antibodies

“…A total of 108 of 362 sputum specimens processed were positive for mycobacteria (acid-fast bacilli). Fifty-one cultures tested positive with MPB64-ICA, and the presence of the M. tuberculosis complex in these cultures was demonstrated by other confirmation tests (Table 4) It is well known that M. bovis BCG vaccine strains can be divided into two groups on the basis of several features, such as the number of IS6110 copies (9,25) and the presence of MPB64 antigen (12,21) and methoxymycolates (19). As shown in Table 5, these two groups of the substrains were clearly differentiated by MPB64-ICA.…”

Detection of mycobacterium tuberculosis in clinical specimens by polymerase chain reaction and Gen-Probe Amplified Mycobacterium Tuberculosis Direct Test

“…The number of copies of the IS986 element varies from 1 to 19, although most strains have between 8 and 15 copies (18). The use of these oligonucleotides may be useful for increasing PCR sensitivity.…”

Detection of Mycobacterium tuberculosis in clinical specimens by polymerase chain reaction and Gen-Probe Amplified Mycobacterium Tuberculosis Direct Test

“…Two oligonucleotides based on the sequence of IS986, whose presence is restricted to organisms belonging to the M. tuberculosis complex, were used as primers for PCR. The number of copies of the IS986 element varies from 1 to 19, although most strains have between 8 and 15 copies (18). The use of these oligonucleotides may be useful for increasing PCR sensitivity.…”