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Journal of Clinical Microbiology, May 2000, p. 2018-2020, Vol. 38, No. 5
Laboratoire de Bactériologie,
Virologie, Hygiène1 and Service de
Médecine Interne et de Maladies
Infectieuses,2 Hôpital Robert
Debré, 51092 Reims cedex, and Laboratoire de
Référence des Mycobactéries, Institut Pasteur,
75724 Paris cedex 15,3 France
Received 5 November 1999/Returned for modification 27 December
1999/Accepted 31 January 2000
A nonchromogenic Mycobacterium species was isolated
from an AIDS patient with acute lymphadenitis. On the basis of the
results of conventional tests, the strain appeared to be an atypical
nonphotochromogenic Mycobacterium kansasii strain.
Sequencing of the 16S rRNA gene revealed a unique nucleic acid
sequence, suggesting that the isolate represents an undescribed
pathogenic species.
In December 1993, a 47-year-old human
immunodeficiency virus-infected man was admitted with new-onset left
axillar adenopathy. His past medical history included syphilis,
gonorrhea, and chronic active hepatitis B. The patient had been treated
with zidovudine since 1991, when his CD4+-cell count had
dropped to below 300 × 106/liter. In 1993 he
developed Kaposi's sarcoma and chronic oral candidiasis. Physical
examination revealed a firm, painful left axillar adenopathy (1.5 by
1.5 cm). The patient had no fever, pulmonary signs, or enteric
disorders. The white cell count was 3,800 × 106/liter, with 50% neutrophils and 30% lymphocytes, and
his CD4+-cell count was 24 × 106/liter.
Chest X rays showed no abnormalities. Surgical ablation of the
adenopathy was performed. Histopathology showed granulomatous inflammation with focal noncaseating necrosis and diffuse acid-fast bacilli (>100/field). The excised tissue was cultured for mycobacteria and was inoculated into Middlebrook 7H12 medium (BACTEC 12B; Becton Dickinson Diagnostic Instruments, Sparks, Md.). Therapy consisted of
isoniazid, rifampin, and ethambutol for 3 months, followed by isoniazid
and rifampin for only 2 months because of nausea. The patient died of
Pseudomonas aeruginosa pulmonary infection 4 months after
the therapy was discontinued. At that time there was no new adenopathy,
and blood and sputum remained negative for acid-fast bacilli.
Microbiological investigation.
Mycobacterial growth was
detected by the BACTEC radiometric method (Becton Dickinson, Towson,
Md.) after 7 days of incubation. Since culture confirmation testing
with use of a commercially available DNA probe for the
Mycobacterium avium-M. intracellulare complex (Accu-Probe;
Gen Probe Inc., San Diego, Calif.) was negative, identification was
performed by biochemical tests (13), mycolic acid analysis
(13), PCR-restriction fragment length polymorphism analysis
(PRA) of the amplified hsp-65 gene (10), and 16S
rRNA gene sequencing (7). The isolate grew as smooth,
nonphotochromogenic colonies on Lowenstein-Jensen medium within 28 days
at temperatures of 22 and 37°C but failed to grow at 41°C. Colonies
grew on thiophen-2 carboxylic acid hydrazide and
p-nitrobenzoate but not on hydroxylamine or NaCl (5%). The
test for niacin production was negative, and the tests for heat-stable
catalase, nitrate reductase, Tween hydrolysis, and arylsulfatase were
positive (Table 1). The strain was
susceptible to rifampin, ethambutol, and pyrazinamide and was resistant
to isoniazid and streptomycin as determined with a radiometric system (Becton Dickinson). The mycolate pattern was I, III, and IV. According to these phenotypic data, the strain was considered most like an
unusual M. kansasii strain with no photochromogenic ability. However, the molecular markers did not confirm this identification. The
polymorphism of the hsp-65 gene, which encodes the 65-kDa heat shock protein, was investigated by PRA. The PRA pattern matched the M. triplex (1) and M. lentiflavum
(8) PRA patterns. The 16S rRNA gene sequence within
hypervariable region A was unique and differed from all previously
published mycobacterial sequences in the EMBL nucleotide sequence
database. The closest sequence was that of M. interjectum
(6), with two nucleotide differences (Fig.
1). The sequences of the hypervariable
regions A of M. kansasii, M. triplex, and
M. lentiflavum showed six, six, and four different nucleotides, respectively (Fig. 1). It must be noted that the PRA
pattern of our strain was distinct from the PRA pattern of M. interjectum. The combined results of the phenotypic and genotypic analyses indicate that our strain does not belong to any described Mycobacterium species.
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Isolation of an Unusual Mycobacterium
Species from an AIDS Patient with Acute Lymphadenitis
ABSTRACT
Top
Abstract
Case Report
References
CASE REPORT
Top
Abstract
Case Report
References
TABLE 1.
Comparison of conventional biochemical and cultural
results, mycolate pattern, and susceptibility pattern for our
isolate and some of the more related speciesa
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FIG. 1.
Partial alignment within hypervariable region A of
selected mycobacterial 16S rRNA sequences. The sequence from M. tuberculosis was used as the reference sequence. Only nucleotides
different from those in the M. tuberculosis sequence are
shown. The first nucleotide corresponds to Escherichia coli
position 40 or M. tuberculosis position 129 (7).
Asterisks indicate nucleotide differences within the sequences of
M. tuberculosis and our strain (second row) and within the
sequences of our strain and any other species included in the
comparative analysis (last row).
Discussion. Previous reports have already described disagreements between phenotypic and genotypic features (11, 12), suggesting that the strains were new Mycobacterium species. During routine application of 16S rRNA sequence determination for the identification of mycobacteria, it has become clear that the genus Mycobacterium is much more diverse than was previously anticipated and harbors a significant number of yet to be described pathogens. Lymphadenitis represents the most frequent extrapulmonary disease due to mycobacteria other than tubercle bacillus infection in children (2, 5). Recently, the incidence of the disease appeared to be increasing, along with a shift in the spectrum of mycobacterial species involved. Since 1996 the list of species that cause lymphadenitis was extended with the description of M. lentiflavum, M. triplex, M. heidelbergense, and M. interjectum (1, 3, 4, 9). In all reported cases, infections due to these agents often occurred in the cervical lymph nodes of children. The pathogenic role of our strain was strongly supported by the site of isolation, the presence of acid-fast bacilli on direct examination, and histopathological observation. The clinical case reported here shows that other mycobacterial species that lack a former description may cause lymphadenitis in AIDS patients.
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ACKNOWLEDGMENTS |
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We thank A. Varnerot for sequencing analysis.
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FOOTNOTES |
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* Corresponding author. Mailing address: Laboratoire de Bactériologie, Virologie, Hygiène, Hôpital Robert Debré, avenue du général Koenig, 51092 Reims cedex, France. Phone: 33-326783993. Fax: 33-326865197. E-mail: obajolet{at}chu-reims.fr.
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REFERENCES |
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Abstract
Case Report
References
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Journal of Clinical Microbiology, May 2000, p. 2018-2020, Vol. 38, No. 5
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
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