Molecular detection of Treponema pallidum ssp. pallidum in clinical samples: sequencing of TP0136, TP0548 and 23S rRNA genes.
|Year of publication||2011|
|MU Faculty or unit|
|Description||Syphilis, caused by the spirochete Treponema pallidum subsp. pallidum (TPA), is a sexually transmitted infectious disease with worldwide occurrence. During the years 2004 - 2010, 119 PCR positive specimens from 91 patients (76 males with median age of 33 years and 15 females with median age of 28 years) were collected in the Czech Republic. Treponemal DNA was detected in 4 types of clinical material: genitoanal and pharyngeal swabs (76 samples), whole blood samples (41), cerebrospinal fluid (1) and blood serum (1), especially in patients with primary (60 patients) and secondary (20 patients) stage of syphilis. Primary screening of clinical specimens included nested PCR detection of two TPA specific loci (tmpC and polA genes). To further characterize the treponemal strains, two sequentially variable genes including TP0136 and TP0548 were amplified and sequenced together with the 23S rRNA gene. Altogether, 64.7% of TPA strains were completely typed and 9 treponemal subtypes were identified among 91 tested patients. Mutations in the 23S rRNA gene (A2058G and A2059G), causing macrolide resistance of TPA strains, were found in samples taken from 28.6% of patients. 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.|