Detection by polymerase chain reaction of Treponema pallidum DNA in cerebrospinal fluid from neurosyphilis patients before and after antibiotic treatment

Noordhoek, G. T.; Wolters, Erik Ch.; Jonge, M. E. J. De; Embden, J. D. A. Van

doi:10.1128/jcm.29.9.1976-1984.1991

Cited by 128 publications

(46 citation statements)

References 40 publications

(40 reference statements)

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“…The sensitivity of our PCR depended on the solvent of the target DNA, since the sensitivity decreased 10 times when artificially seeded CSF or urine samples were used compared with the sensitivity when a spirochetal solution in PBS was used. This phenomenon was described previously (8,40) and is most likely due to Taq polymerase inhibitors in body fluids. Results of our experiment with different amounts of target DNA preparation and the double amplification of B. burgdorferi DNA and the human gastrin gene in parallel illustrate this problem (Fig.…”

Section: Discussionsupporting

confidence: 64%

“…The diagnostic sensitivity achieved in patients with this condition seems higher than that achieved in patients with neuroborreliosis. This and the sporadically positive PCR results obtained months to years after therapy in a few patients with neurosyphilis (40) may be attributable to the significantly higher number of spirochetes in patients with syphilis compared with the number in those with Lyme borreliosis.…”

Section: Discussionmentioning

confidence: 99%

“…Recently, T. pallidum DNA has successfully been detected in CSF from patients with neurosyphilis (8,15,24,40). The diagnostic sensitivity achieved in patients with this condition seems higher than that achieved in patients with neuroborreliosis.…”

Section: Discussionmentioning

confidence: 99%

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Detection of Borrelia burgdorferi DNA in urine samples and cerebrospinal fluid samples from patients with early and late Lyme neuroborreliosis by polymerase chain reaction

1992

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A polymerase chain reaction (PCR) was developed for use in the identification of a 248-bp fragment of the Borrelia burgdorferi flagellin gene in urine and cerebrospinal fluid (CSF) from patients with Lyme neuroborreliosis. The specificities of the PCR products were confirmed by DNA-DNA hybridization with an internal probe. The assay had a detection limit of 10 in vitro-cultivated B. burgdorferi. The PCR assay seemed to be species wide as well as species specific, since DNA from all 21 B. burgdorferi isolates from humans tested but not from Borrelia hermsii or Treponema pallidum could be amplified. We tested 10 consecutively diagnosed patients with untreated neuroborreliosis. There was lymphocytic pleocytosis and intrathecal B. burgdorferispecific antibody synthesis in the CSF of all patients. Urine and CSF samples were investigated by PCR before, during, and up to 8.5 months after therapy. B. burgdorferi DNA was detected in urine samples from nine patients; five patients, including two patients with chronic neuroborreliosis, were PCR positive prior to treatment, whereas urine samples from the remaining four patients obtained 3 to 6 days after the onset of therapy became PCR positive. All urine samples obtained >4 weeks after therapy were negative by PCR. PCR of CSF was less sensitive, and samples from only four patients, including one with chronic neuroborreliosis, were positive. We conclude that urine is a more suitable sample source than CSF for use in B. burgdorferi DNA detection by PCR. Normalization of inflammatory CSF changes and the negative PCR results during follow-up even in patients with chronic neuroborreliosis do not point to a persistent infection. The future role of PCR as a diagnostic tool for Lyme neuroborreliosis is still uncertain.

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Section: Discussionsupporting

confidence: 64%

Section: Discussionmentioning

confidence: 99%

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Detection of Borrelia burgdorferi DNA in urine samples and cerebrospinal fluid samples from patients with early and late Lyme neuroborreliosis by polymerase chain reaction

1992

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“…Moreover, the finding that microbial DNA can be detected even after successful antimicrobial treatment suggests that the assays detect both viable and non-viable organisms. 119,120 Clearly, interpretive guidelines based on the correlation of test results with clinical presentation and existing standards will be required before these assays can be used for definitive diagnosis and/or treatment decisions.…”

Section: Clinical Significance Of Positive Pcrmentioning

confidence: 99%

PCR-based diagnostics for infectious diseases: uses, limitations, and future applications in acute-care settings

Yang

Rothman

2004

The Lancet Infectious Diseases

796

545

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Molecular diagnostics are revolutionising the clinical practice of infectious disease. Their effects will be significant in acute-care settings where timely and accurate diagnostic tools are critical for patient treatment decisions and outcomes. PCR is the most well-developed molecular technique up to now, and has a wide range of already fulfilled, and potential, clinical applications, including specific or broad-spectrum pathogen detection, evaluation of emerging novel infections, surveillance, early detection of biothreat agents, and antimicrobial resistance profiling. PCR-based methods may also be cost effective relative to traditional testing procedures. Further advancement of technology is needed to improve automation, optimise detection sensitivity and specificity, and expand the capacity to detect multiple targets simultaneously (multiplexing). This review provides an up-to-date look at the general principles, diagnostic value, and limitations of the most current PCR-based platforms as they evolve from bench to bedside.

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“…These methodologies, particularly the PCR, have been used in the detection of treponemes. This technique has detected T. pallidum in a variety of clinical specimens, including serum (1,3,18), cerebrospinal fluid (1,3,5,12,18), amniotic fluid (1,3,18), fixed tissues (1), and lesion exudate (8,13,22). However, its performance relative to other tests, particularly in early disease, remains to be validated by extensive clinical studies.…”

mentioning

confidence: 99%

Comparison of molecular and microscopic techniques for detection of Treponema pallidum in genital ulcers

et al. 1995

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We compared the ability of direct immunofluorescent staining (DFA) and the PCR to detect Treponema pallidum in specimens from patients with genital ulcer disease. Touch preparations from 156 patients with genital lesions were fixed in acetone and stained with a fluorescein-labeled monoclonal antibody specific for the 37-kDa antigen of T. pallidum. After microscopic examination, the smear was removed from the slide with a swab. DNA was extracted with phenol-chloroform and precipitated with isopropanol. Ten microliters of the extracted DNA was amplified by PCR using primers for the gene encoding the 47-kDa protein of T. pallidum and hybridized to an internal probe. Twenty-two of 156 specimens were positive for T. pallidum by DFA and PCR, while 127 were negative by both methods, yielding a concordance of 95.5% ( ‫؍‬ 0.84). Four specimens were positive by PCR and negative by DFA, while three specimens were negative by PCR and positive by DFA. The DFA-negative, PCR-positive specimens may have resulted from the presence of large numbers of leukocytes on the slides, obscuring visualization of treponemes. The DFA-positive, PCR-negative results were not due to inhibition of the PCR since purified T. pallidum DNA was amplified when added to aliquots of these specimens. Negative results in these specimens were most likely due to inefficient recovery of their DNA. These data suggest that DFA and PCR are equivalent methods for detection of T. pallidum on touch preparations of genital lesions. Further refinements of the PCR assay are necessary for it to significantly improve the detection of T. pallidum in genital lesions.

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Detection by polymerase chain reaction of Treponema pallidum DNA in cerebrospinal fluid from neurosyphilis patients before and after antibiotic treatment

Cited by 128 publications

References 40 publications

Detection of Borrelia burgdorferi DNA in urine samples and cerebrospinal fluid samples from patients with early and late Lyme neuroborreliosis by polymerase chain reaction

Detection of Borrelia burgdorferi DNA in urine samples and cerebrospinal fluid samples from patients with early and late Lyme neuroborreliosis by polymerase chain reaction

PCR-based diagnostics for infectious diseases: uses, limitations, and future applications in acute-care settings

Comparison of molecular and microscopic techniques for detection of Treponema pallidum in genital ulcers

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