This is the first identified case of Mycobacterium bovis bacillus Calmette-Guérin (BCG)-derived cutaneous tuberculosis that localizes at a place different from the vaccination site in hosts without immune deficiency. A healthy baby with a developing abscess is described. A multiplex PCR identified the abscess as originating from M. bovis BCG Tokyo 172. CASE REPORTA 6-month-old girl with no previous significant medical problems developed an abscess in her left humerus. She had been vaccinated with Mycobacterium bovis bacillus CalmetteGuérin (BCG) Tokyo 172 at the age of 4 months. At 5 months, a red and swelling nodule appeared in her left humerus at a place different from the vaccination site. At 6 months, she visited our hospital. A red, fluctuating, and swollen abscess about 2 cm from the region of the vaccination was found. The abscess measured 10 by 15 mm. We found no swelling of the lymph nodes. The patient was afebrile and did not have respiratory symptoms. There was no abnormal chest radiographic finding. Skin testing with purified protein derivative tuberculin was positive with 10-by 7-mm erythema, without an induration or surrounding erythema. There was no family history of a similar reaction to BCG vaccination. The history of the girl's grandmother revealed pulmonary tuberculosis in her twenties. In addition, the possibility of close contact with other tuberculosis patients could not be excluded.We punctured the abscess and submitted the pus for conventional microbial culture, mycobacterial culture, amplification of mycobacterial nucleic acid using an AMPLICOR MTB microwell plate assay kit (Roche Diagnostic Systems, Branchburg, N.J.), and microscopic examination of a smear. Conventional microbial culture failed to detect any significant pathogens. The smear examination was negative, but amplification of nucleic acid using a primer for a Mycobacterium tuberculosis complex-specific region of the 16S rRNA gene (KY172-T3) was positive for M. tuberculosis complexes. A 4-week mycobacterial culture grew an M. tuberculosis complex colony. This colony showed sensitivity to isoniazid, rifampin, ethambutol, and streptomycin and resistance to pyrazinamide, suggesting that the colony may have derived from BCG Tokyo 172. However, we could not clearly distinguish the origin among M. tuberculosis complex strains (M. tuberculosis, M. bovis BCG, or other strains of M. bovis).For the purpose of quick and accurate identification of the sample, we ran multiplex PCR on the sample by a simple modification of the method of Bedwell et al. (1). In 1999, a DNA microarray clarified that 16 regions in BCG strains, named RD1 to RD16, were different from those in M. tuberculosis H37Rv and that some regions differed among BCG strains (2). RD1 is not present in all M. bovis BCG strains, but it is present in other strains of the M. tuberculosis complex. For M. bovis BCG strains, Talbot et al. designed primers to amplify the complement sequence of RD1 (9). Another PCR procedure based on the intergenic region named SenX3-RegX3 was reporte...
The genomic DNA fragment which contains ribosomal RNA (rRNA) genes for Treponema phagedenis was cloned into bacteriophage vector lambda EMBL3. A restriction map of the fragment was constructed and the organization of the rRNA genes was determined. The fragment contained at least one copy of the 16S, 23S and 5S sequences and the genes are arranged in the order 16S-23S-5S . Southern hybridization using radiolabeled rRNA gene probes to genomic DNA from T . phagedenis strain Reiter and T. pallidum strain Nichols showed that these organisms have two radioactive fragments which hybridize to the probes in their genome . These results suggest that both pathogenic and non-pathogenic strains of Treponema may carry at least two sets of rRNA genes on their chromosomes.We are interested in studying the organization and regulation of the genes coding for the ribosomal RNAs (rRNA) of spirochetes. Our interest in rRNA genes in spirochetes has arisen because the organization of rRNA genes for leptospires is unique (5). In addition, our finding of a functional promoter immediately preceding one operon comprising only the 5S rRNA gene gave further evidence that the other rRNA genes of the strains of leptospires are organized in separate transcription units. By the transcriptional analysis of the 5' regions of the 16S and 23S rRNA genes for Leptospira interrogans, primary transcripts were detected and the sequence resembling a promoter was seen at upstream from the 5'-terminus of the gene in each case.Organization and transcription of genes for rRNA have been highly conserved among procaryotes because the synthesis of rRNA is an essential process in the growth of any organism (13,14). In most eubacterial species so far examined, the sequences for the three rRNAs are closely linked in gene sets , in the order 16S-23S-5S (11). Some exceptions to this rule have been reported . In Thermus thermophilus (21), 16S and 23S rRNA genes are separated; in Mycoplasma gallisepticum (2), the 16S gene is separated from the other genes; and in another Mycoplasma strain (20), the 161
The intravesical instillation of bacillus Calmette-Guérin (BCG) is a standard therapy for superficial bladder carcinoma. Tuberculosis-like inflammation in the genitourinary tract is a serious complication of BCG. It can occur after a long interval from the cessation of the intravesical BCG therapy. If inflammation occurs, it is necessary to test whether the BCG strain has caused it or another mycobacterium species has. However, there has never been a report that proves BCG causes the inflammation, because BCG is difficult to differentiate from other strains of Mycobacterium bovis and other members of the Mycobacterium tuberculosis complex by conventional tests, including regular polymerase chain reaction (PCR). We first present a case of epididymo-orchitis, which developed 31 months after the cessation of BCG therapy, detected using a multiplex PCR method as having been caused by BCG. Our report illustrates the efficacy of this method to detect the responsible microbe that is thought to be transmitted from the instillated BCG strain.
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