Previous Article
Antimicrobial Agents and Chemotherapy, March 2000, p. 806-807, Vol. 44, No. 3
0066-4804/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
LETTERS TO THE EDITOR
A Point Mutation Associated with Bacterial Macrolide Resistance
Is Present in Both 23S rRNA Genes of an Erythromycin-Resistant
Treponema pallidum Clinical Isolate
 |
LETTER |
Treponema pallidum subsp. pallidum (T. pallidum) is the noncultivable agent of syphilis, a sexually
transmitted disease that is a risk factor for HIV infection
(9). Penicillin is the preferred drug for treatment of
syphilis (1). Alternative antibiotics for nonpregnant
penicillin-allergic patients with primary or secondary syphilis include
doxycycline, tetracycline, or erythromycin. Erythromycin treatment
failures have been documented in patients with primary or secondary
syphilis and infants with congenital syphilis (2, 3, 5). We
previously demonstrated high-level erythromycin resistance
(Ermr) in a T. pallidum clinical isolate (Street
strain 14) obtained from a syphilis patient who failed erythromycin
therapy (11). Macrolide resistance usually results from
target site alteration due to methylation or mutations in the
peptidyltransferase region of 23S rRNA. Mutations of the adenine (A)
residue cognate to position A2058 or A2059 in the Escherichia
coli 23S rRNA gene are associated with macrolide resistance in
bacteria that contain one or two copies of these genes (6-8,
13). This observation prompted us to analyze the corresponding
regions of the two 23S rRNA genes of T. pallidum Street
strain 14 (Ermr); T. pallidum Nichols strain, an
erythromycin-sensitive (Erms) control (11); and
the closely related yaws agent, T. pallidum subsp.
pertenue (T. pertenue) Gauthier strain
(Erms) (11).
T. pallidum strains were grown by testicular cultivation in
rabbits, and genomic DNA was extracted as previously described (11, 12). T. pertenue genomic DNA was provided by
A. Centurion-Lara and S. Lukehart. Both copies of the 23S rRNA genes
were PCR amplified from treponemal genomic DNA (Expand Long Template
PCR System; Boehringer-Mannheim, Indianapolis, Ind.). Oligonucleotide
primers were designed based on sequence data from the T. pallidum Nichols genome sequencing project (GenBank accession no.
AE001204 and AE001208) (4). Gel-purified PCR amplicons were
cloned (pGEM-T Easy vector; Promega Corp., Madison, Wis.), and both
strands of the inserts were sequenced at the University of North
Carolina, Chapel Hill, Automated DNA Sequencing Facility as previously
described (12).
A 692-bp region (representing 133 nucleotides 5' and 558 nucleotides 3'
of the position cognate to A2058 in the E. coli 23S rRNA
gene) was analyzed for T. pallidum and T. pertenue. The nucleotide sequences of this region of both T. pallidum Nichols strain 23S rRNA genes were identical to those of
the Nichols strain in GenBank (4). Additionally, the
sequences of the same 692-bp region of both T. pertenue
Gauthier strain 23S rRNA genes were identical to those of the T. pallidum Nichols strain. Interestingly, the sequences of the
692-bp region of both T. pallidum Street strain 14 23S rRNA
genes were identical to those of T. pallidum Nichols strain
except that the Street strain 14 sequences contained a guanine (G) at
the position cognate to A2058 (Table 1).
The A-to-G transition mutation was present in all clones sequenced for
both Street strain 14 23S rRNA genes. The identical mutation was also present when the Street strain 14 23S rRNA genes were PCR amplified and
directly sequenced.
View this table:
[in this window]
[in a new window]
|
TABLE 1.
Comparison of the 23S rRNA gene sequences of T. pallidum Nichols and Street strain 14 and T. pertenue
in the region containing the cognate to E. coli rDNA A2058
|
|
Since rRNA methylase genes do not appear to be present in T. pallidum Street strain 14 (L. Stamm, unpublished data), we propose that the A-to-G transition mutations in both 23S rRNA genes of this
spirochete are responsible for its high-level resistance to
erythromycin and related macrolides (roxithromycin
[11] and azithromycin [10]). Similar
point mutations in the 23S rRNA genes of other T. pallidum
Street strains may account for erythromycin treatment failures observed
in syphilis patients who have complied with their therapeutic regimen.
Our results also demonstrate the utility of the T. pallidum
Nichols strain complete genome sequence for investigating the genetic
basis of antimicrobial resistance in T. pallidum.
Nucleotide sequence accession numbers.
The sequences of the
692-bp region of the 23S rRNA genes of T. pallidum Nichols
strain and Street strain 14 and T. pertenue Gauthier strain
have been deposited in GenBank under accession numbers AF200367,
AF200365, and AF200366, respectively.
| |
ACKNOWLEDGMENTS |
This research was supported by National Institutes of Health grant AI31496.
| |
FOOTNOTES |
*
Phone: (919) 966-3882
Fax: (919) 966-2089
E-mail: Istamm{at}email.unc.ed
| |
REFERENCES |
| 1.
|
Centers for Disease Control and Prevention.
1998.
Guidelines for treatment of sexually transmitted diseases.
Morbid. Mortal. Weekly Rep.
47(RR-1):28-48.
|
| 2.
|
Duncan, W. C.
1989.
Failure of erythromycin to cure secondary syphilis in a patient infected with the human immunodeficiency virus.
Arch. Dermatol.
125:82-84[Abstract/Free Full Text].
|
| 3.
|
Fenton, L. J., and I. J. Light.
1976.
Congenital syphilis after maternal treatment with erythromycin.
Obstet. Gynecol.
47:492-494[Medline].
|
| 4.
|
Fraser, C. M.,
S. J. Norris,
G. M. Weinstock,
O. White,
G. G. Sutton,
R. Dodson,
M. Gwinn,
E. K. Hickey,
R. Clayton,
K. A. Ketchum,
E. Sodergren,
J. M. Hardham,
M. P. McLeod,
S. Salzberg,
J. Peterson,
H. Khalak,
D. Richardson,
J. K. Howell,
M. Chidambaram,
T. Utterback,
L. McDonald,
P. Artiach,
C. Bowman,
M. D. Cotton,
C. Fujii,
S. Garland,
B. Hatch,
K. Horst,
K. Roberts,
M. Sandusky,
J. Weidman,
H. O. Smith, and J. C. Venter.
1998.
Complete genome sequence of Treponema pallidum, the syphilis spirochete.
Science
281:375-388[Abstract/Free Full Text].
|
| 5.
|
Hashisaki, P.,
G. G. Wertzberger,
G. L. Conrad, and C. R. Nichols.
1983.
Erythromycin failure in treatment of syphilis in a pregnant woman.
Sex. Transm. Dis.
10:36-38[CrossRef][Medline].
|
| 6.
|
Karlsson, M.,
C. Fellstrom,
M. U. K. Heldtander,
K.-E. Johansson, and A. Franklin.
1999.
Genetic basis of macrolide and lincosamide resistance in Brachyspira (Serpulina) hyodysenteriae.
FEMS Microbiol. Lett.
172:255-260[CrossRef][Medline].
|
| 7.
|
Lucier, T. S.,
K. Heitzman,
S. K. Liu, and P. C. Hu.
1995.
Transition mutations in the 23S rRNA of erythromycin-resistant isolates of Mycoplasma pneumoniae.
Antimicrob. Agents Chemother.
39:2770-2773[Abstract].
|
| 8.
|
Sander, P.,
T. Prammananan,
A. Meier,
K. Frischkorn, and E. C. Bottger.
1997.
The role of ribosomal RNAs in macrolide resistance.
Mol. Microbiol.
26:469-480[CrossRef][Medline].
|
| 9.
|
Stamm, L. V.
1999.
Biology of Treponema pallidum, p. 467-472.
In
K. K. Holmes, P. F. Sparling, P.-A. Mardh, S. M. Lemon, W. E. Stamm, P. Piot, and J. M. Wasserheit (ed.), Sexually transmitted diseases. McGraw-Hill, New York, N.Y.
|
| 10.
|
Stamm, L. V., and E. A. Parrish.
1990.
In-vitro activity of azithromycin and CP-63,956 against Treponema pallidum.
J. Antimicrob. Chemother.
25(Suppl. A):11-14.
|
| 11.
|
Stamm, L. V.,
J. T. Stapleton, and P. J. Bassford, Jr.
1988.
In vitro assay to demonstrate high-level erythromycin resistance of a clinical isolate of Treponema pallidum.
Antimicrob. Agents Chemother.
32:164-169[Abstract/Free Full Text].
|
| 12.
|
Stamm, L. V.,
S. R. Greene,
H. L. Bergen,
J. M. Hardham, and N. Y. Barnes.
1998.
Identification and sequence analysis of Treponema pallidum tprJ, a member of a polymorphic multigene family.
FEMS Microbiol. Lett.
169:155-163[CrossRef][Medline].
|
| 13.
|
Taylor, D. E.,
Z. Ge,
D. Purych,
T. Lo, and K. Hiratsuka.
1997.
Cloning and sequence analysis of two copies of a 23S rRNA gene from Helicobacter pylori and association of clarithromycin resistance with 23S rRNA mutations.
Antimicrob. Agents Chemother.
41:2621-2628[Abstract].
|
| | | | |
L. V. Stamm*
H. L. Bergen
Program in Infectious Diseases
Department of Epidemiology
School of Public Health
The University of North Carolina
Chapel Hill, North Carolina 27599-7400
|
Antimicrobial Agents and Chemotherapy, March 2000, p. 806-807, Vol. 44, No. 3
0066-4804/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
This article has been cited by other articles:
-
Matejkova, P., Flasarova, M., Zakoucka, H., Borek, M., Kremenova, S., Arenberger, P., Woznicova, V., Weinstock, G. M., Smajs, D.
(2009). Macrolide treatment failure in a case of secondary syphilis: a novel A2059G mutation in the 23S rRNA gene of Treponema pallidum subsp. pallidum. J Med Microbiol
58: 832-836
[Abstract]
[Full Text]
-
Pandori, M. W., Gordones, C., Castro, L., Engelman, J., Siedner, M., Lukehart, S., Klausner, J.
(2007). Detection of Azithromycin Resistance in Treponema pallidum by Real-Time PCR. Antimicrob. Agents Chemother.
51: 3425-3430
[Abstract]
[Full Text]
-
LaFond, R. E., Lukehart, S. A.
(2006). Biological Basis for Syphilis. Clin. Microbiol. Rev.
19: 29-49
[Abstract]
[Full Text]
-
Lukehart, S. A., Godornes, C., Molini, B. J., Sonnett, P., Hopkins, S., Mulcahy, F., Engelman, J., Mitchell, S. J., Rompalo, A. M., Marra, C. M., Klausner, J. D.
(2004). Macrolide Resistance in Treponema pallidum in the United States and Ireland. NEJM
351: 154-158
[Full Text]
-
Stamm, L. V., Bergen, H. L., Shangraw, K. A.
(2001). Natural Rifampin Resistance in Treponema spp. Correlates with Presence of N531 in RpoB Rif Cluster I. Antimicrob. Agents Chemother.
45: 2973-2974
[Full Text]
-
Tait-Kamradt, A., Davies, T., Appelbaum, P. C., Depardieu, F., Courvalin, P., Petitpas, J., Wondrack, L., Walker, A., Jacobs, M. R., Sutcliffe, J.
(2000). Two New Mechanisms of Macrolide Resistance in Clinical Strains of Streptococcus pneumoniae from Eastern Europe and North America. Antimicrob. Agents Chemother.
44: 3395-3401
[Abstract]
[Full Text]
