Molecular detection of Treponema pallidum in secondary and tertiary syphilis

Treponema pallidum can be detected by conventional techniques such as dark-field microscopy, immunofluorescence or the rabbit infectivity test, in large numbers in the skin lesions of primary and early secondary syphilis. In the skin lesions of late secondary and tertiary syphilis, conventional techniques fail to detect spirochaetes in general, perhaps due to increasing degeneration and the disappearance of treponemal spirochaetes in late syphilitic skin lesions. We used the highly sensitive technique of polymerase chain reaction (PCR) to prove the presence of Treponema pallidum-specific DNA in six lesions of late secondary syphilis and seven lesions of tertiary syphilis, including one syphilitic gumma, A Whartin-Starry stain was carried out in all 13 specimens and did not reveal any treponemal structures. Treponema pallidum-specific DNA was amplified by PCR in four of six cases of secondary syphilis and in the syphilitic gumma. These results are in favour of a direct cell-mediated immune reaction directed against treponemal antigen rather than the concept of an Id-reaction. Beside the usefulness of a PCR-based assay for understanding the aetiology of lesions of late syphilis, the assay described can be of clinical importance in various situations where traditional methods fail to detect Treponema pallidum because of lack of sensitivity

Clinical and molecular assessment of ceftriaxone therapy in acrodermatitis chronica atrophicans

To assess the presence of Borrelia burgdorferi (Bb)-specific DNA in acrodermatitis chronica atrophicans (ACA) lesions before and after ceftriaxone (CE) therapy by polymerase chain reaction (PCR). Biopsy specimens of seven patients (m:f = 0:7, mean age 59 yrs) with an inflammatory stage of ACA (bluish-red discoloration and swelling) along with slight signs of skin atrophy before and immediately after CE therapy (1×2g i.V., 14 days) have been studied. PCR analysis from formalin-fixed, paraffin-embedded material has been performed as described (Wienecke R et al., 1993). In pre-treatment specimens, Bb-specific DNA was amplified by PCR in 5/7 ACA patients (71%). After CE therapy, PCR yielded negative results in all specimens. Clinically, no more swelling could be observed directly after therapy, whereas erythema and atrophy remained nearly unchanged. Ten weeks after therapy, only a frail bluish-red discoloration and atrophy were present, but skin had still not turned to normal in any of the ACA patients. The slow improvement of inflammatory lesions, and the persistence of skin atrophy, as found in our patients, is well known from the literature, also following treatment with various other antibiotics than CE, and does not necessarily indicate a treatment failure (Weber K et al., 1988). Our PCR data further substantiate this fact by demonstrating the eradication of Bb from the site of infection by the CE regimen performed. However, it remains to be determined, if a longer treatment reqimen leads to a better clinical response

Polymerase chain reaction control of antibiotic treatment in dermatoborreliosis

Assessment of the efficacy of an antibiotic drug used in patients with various manifestations of dermatoborreliosis is crucial, Clinical judgement alone (resolution of the present dermatologic lesion, prevention of later major or minor sequelae) is not sufficient in erythema migrans and acrodermatitis chronica atrophicans, Thus, laboratory tests are desirable to prove the benefit of an antimicrobial agent, It was intended to establish a constant parameter - besides the clinical picture - for assessing the efficacy of antibiotic treatment in patients with dermatoborreliosis in terms of eradication of Borrelia burgdorferi from the site of infection, Polymerase chain reaction (PCR) was therefore performed from pretreatment biopsy specimens from lesional skin of 36 erythema migrans patients (m:f = 15:21, mean age 49 years) and seven acrodermatitis chronica atrophicans patients (m:f = 0:7, mean age 59 years), respectively, After antibiotic therapy with minocycline (100 mg, orally twice daily, 14 days) for erythema migrans, and ceftriaxone (2 g, intravenously once daily, 14 days) for acrodermatitis chronica atrophicans another punch biopsy was obtained and analysed by PCR, In pretreatment specimens, B, burgdorferi-specific DNA was amplified by PCR in 23/36 erythema migrans patients (69%), and in 5/7 acrodermatitis chronica atrophicans patients (71%), After antibiotic therapy, PCR yielded negative results in all of these cases, Clinically all patients showed complete recovery or at least marked improvement of lesions at this time, PCR appears to be a reliable parameter for the assessment of the efficacy of antibiotic treatment in dermatoborreliosis