Molecular detection and typing of Treponema pallidum ssp.
pallidum in clinical samples based on sequencing of TP0136,
TP0548 and 23S rRNA genes

a 2011

Molecular detection and typing of Treponema pallidum ssp. pallidum in clinical samples based on sequencing of TP0136, TP0548 and 23S rRNA genes

MIKALOVÁ, Lenka, Petra POSPÍŠILOVÁ, Magdalena FLASAROVÁ, Zuzana VALIŠOVÁ, Vladana WOZNICOVÁ et. al.

Basic information

Original name

Molecular detection and typing of Treponema pallidum ssp. pallidum in clinical samples based on sequencing of TP0136, TP0548 and 23S rRNA genes

Authors

MIKALOVÁ, Lenka (203 Czech Republic, guarantor, belonging to the institution), Petra POSPÍŠILOVÁ (203 Czech Republic, belonging to the institution), Magdalena FLASAROVÁ (203 Czech Republic, belonging to the institution), Zuzana VALIŠOVÁ (203 Czech Republic, belonging to the institution), Vladana WOZNICOVÁ (203 Czech Republic, belonging to the institution), H. ZÁKOUCKÁ (203 Czech Republic), I. KUKLOVÁ (203 Czech Republic) and David ŠMAJS (203 Czech Republic, belonging to the institution)

Edition

Acta Microbiologica et Immunologica Hungarica, vol. 58, Supplement : 16th International Congress of the Hungarian Society for Microbiology, 2011

Other information

Language

English

Type of outcome

Konferenční abstrakt

Field of Study

10600 1.6 Biological sciences

Country of publisher

Hungary

Confidentiality degree

není předmětem státního či obchodního tajemství

Impact factor

Impact factor: 0.787

RIV identification code

RIV/00216224:14110/11:00050392

Organization unit

Faculty of Medicine

ISSN

Keywords in English

Treponema pallidum

Změněno: 18/1/2012 14:54, Mgr. Michal Petr

Abstract

V originále

Syphilis, caused by the spirochete Treponema pallidum subsp. pallidum (TPA), is a sexually transmitted infectious disease with worldwide occurrence. The bacterium TPA cannot be cultivated in vitro, therefore laboratory diagnostics is traditionally made by microscopy and serology testing, which do not provide epidemiological data. Molecular detection of treponemes could type the clinical samples and provide the antibiotic susceptibility information. During the years 2004-2010, 294 patients (415 samples) were tested for the presence of treponemal DNA. Primary screening of clinical specimens included nested PCR detection of two TPA specific loci (tmpC and polA genes). Out of 294 patients, 91 patients were PCR positive. PCR positive patients were more often in the primary stage of syphilis (p=0.0003) compared to the control (PCR negative) group of syphilis seropositive patients. Treponemal DNA was detected in 4 types of clinical material: genitoanal, pharyngeal and skin swabs (75 samples), whole blood samples (42), cerebrospinal fluid (1) and blood serum (1). Molecular typing method was based on amplification and sequencing of two sequentially variable genes including TP0136 and TP0548 together with the 23S rRNA gene, where the mutation in position A2058G or A2059G cause the macrolide resistance. Out of 91 patients with PCR positive samples, 49 patients were completely typed (sequences of TP0136, TP0548 and 23S rRNA genes were determined), 15 patients were partially typed (only two out of 3 loci were detected). Nine different genotypes among the 64 completely or partially typed patients were found. 35.6% of treponemal strains were resistant to macrolide antibiotics. Identified subtypes of TPA strains were further typed with the CDC typing system for TPA treponemes (comprising analysis of the arp and tpr genes). The obtained unique TP0136 and TP0548 sequences were found to combine independently with CDC subtypes indicating their potential for more detailed genetic characterization of TPA-containing clinical samples.

Links

GA310/07/0321, research and development project

Name: Komparativní genomové sekvencování patogenních treponem a transkriptomová analýza T. pallidum ssp. pallidum

Investor: Czech Science Foundation

MSM0021622415, plan (intention)

Name: Molekulární podstata buněčných a tkáňových regulací

Investor: Ministry of Education, Youth and Sports of the CR, Molecular basis of cell and tissue regulations

NT11159, research and development project

Name: Mapování výskytu makrolidové rezistence původce syfilis v ČR a molekulární typizace jednotlivých syfilitických kmenů

Investor: Ministry of Health of the CR

Citovat

MIKALOVÁ, Lenka, Petra POSPÍŠILOVÁ, Magdalena FLASAROVÁ, Zuzana VALIŠOVÁ, Vladana WOZNICOVÁ, H. ZÁKOUCKÁ, I. KUKLOVÁ and David ŠMAJS. Molecular detection and typing of Treponema pallidum ssp. pallidum in clinical samples based on sequencing of TP0136, TP0548 and 23S rRNA genes. In Acta Microbiologica et Immunologica Hungarica, vol. 58, Supplement : 16th International Congress of the Hungarian Society for Microbiology. 2011. ISSN 1217-8950.

@proceedings{966963,
   author = {Mikalová, Lenka and Pospíšilová, Petra and Flasarová, Magdalena and Vališová, Zuzana and Woznicová, Vladana and Zákoucká, H. and Kuklová, I. and Šmajs, David},
   booktitle = {Acta Microbiologica et Immunologica Hungarica, vol. 58, Supplement : 16th International Congress of the Hungarian Society for Microbiology},
   keywords = {Treponema pallidum},
   language = {eng},
   title = {Molecular detection and typing of Treponema pallidum ssp. pallidum in clinical samples based on sequencing of TP0136, TP0548 and 23S rRNA genes},
   year = {2011}
}

TY  - CONF
ID  - 966963
AU  - Mikalová, Lenka - Pospíšilová, Petra - Flasarová, Magdalena - Vališová, Zuzana - Woznicová, Vladana - Zákoucká, H. - Kuklová, I. - Šmajs, David
PY  - 2011
TI  - Molecular detection and typing of Treponema pallidum ssp. pallidum in clinical samples based on sequencing of TP0136, TP0548 and 23S rRNA genes
KW  - Treponema pallidum
N2  - Syphilis, caused by the spirochete Treponema pallidum subsp. pallidum (TPA), is a sexually transmitted infectious disease with worldwide occurrence. The bacterium TPA cannot be cultivated in vitro, therefore laboratory diagnostics is traditionally made by microscopy and serology testing, which do not provide epidemiological data. Molecular detection of treponemes could type the clinical samples and provide the antibiotic susceptibility information. During the years 2004-2010, 294 patients (415 samples) were tested for the presence of treponemal DNA. Primary screening of clinical specimens included nested PCR detection of two TPA specific loci (tmpC and polA genes). Out of 294 patients, 91 patients were PCR positive. PCR positive patients were more often in the primary stage of syphilis (p=0.0003) compared to the control (PCR negative) group of syphilis seropositive patients. Treponemal DNA was detected in 4 types of clinical material: genitoanal, pharyngeal and skin swabs (75 samples), whole blood samples (42), cerebrospinal fluid (1) and blood serum (1). Molecular typing method was based on amplification and sequencing of two sequentially variable genes including TP0136 and TP0548 together with the 23S rRNA gene, where the mutation in position A2058G or A2059G cause the macrolide resistance. Out of 91 patients with PCR positive samples, 49 patients were completely typed (sequences of TP0136, TP0548 and 23S rRNA genes were determined), 15 patients were partially typed (only two out of 3 loci were detected). Nine different genotypes among the 64 completely or partially typed patients were found. 35.6% of treponemal strains were resistant to macrolide antibiotics. Identified subtypes of TPA strains were further typed with the CDC typing system for TPA treponemes (comprising analysis of the arp and tpr genes). The obtained unique TP0136 and TP0548 sequences were found to combine independently with CDC subtypes indicating their potential for more detailed genetic characterization of TPA-containing clinical samples.
ER  -

MIKALOVÁ, Lenka, Petra POSPÍŠILOVÁ, Magdalena FLASAROVÁ, Zuzana VALIŠOVÁ, Vladana WOZNICOVÁ, H. ZÁKOUCKÁ, I. KUKLOVÁ and David ŠMAJS. Molecular detection and typing of Treponema pallidum ssp. pallidum in clinical samples based on sequencing of TP0136, TP0548 and 23S rRNA genes. In \textit{Acta Microbiologica et Immunologica Hungarica, vol. 58, Supplement : 16th International Congress of the Hungarian Society for Microbiology}. 2011. ISSN~1217-8950.

Detailed Information on Publication Record