Detection of Spirochetal DNA from Patients with Neuroborreliosis

Czech version

Authors: D. Pícha; L. Moravcová; D. Smíšková
Authors‘ workplace: Klinika infekčních nemocí 2. LF UK a Nemocnice Na Bulovce, Praha
Published in: Cesk Slov Neurol N 2016; 79/112(5): 566-571
Category: Original Paper

Overview

Aim:
To assess the importance of Borrelia burgdorferi sensu lato DNA verification in neuroborreliosis.

Material and methods:
Seventy-one hospitalized patients with clinically and laboratory-confirmed neuroborreliosis were included. PCR of plasma, urine and CSF was performed in parallel before and after the treatment, and after 3 and 6 months for plasma and urine. DNA was isolated using the QIAamp DNA Mini Kit, PCR was arranged as a two-step amplification (nested PCR). Each sample was examined by the five amplification systems in parallel: two were targeted at plasmide genes encoding OspA and OspC proteins and three at genes for 16S rDNA, flagellin and p66 protein.

Results:
Of all 71 patients, the specific DNA was found in 51 cases (71.8%) before treatment. After the treatment, the DNA was proven in 25 patients (35.2%). PCR positivity persisted in 19 (26.8%) after three months and still in five (7%) patients after six months. The highest frequency of PCR-positive results was obtained by the 16S rDNA system, slightly lower sensitivity was achieved with the OspA, OspC and flagellin primers and the lowest frequency of positives was in the p66 target.

Conclusion:
Borrelial DNA was most frequently found in CSF, less frequently in urine and the lowest number of positive results were in plasma. PCR positivity decreased gradually during the observation period, the test was still positive in some patients after six months. It is possible to recommend testing with a minimum of two target sequences to improve diagnostic sensitivity. Our results suggest that the PCR testing should only be used in neuroborreliosis as an supplementary diagnostic method and subject to correct clinical interpretation.

Key words:
neuroborreliosis – nested PCR – plasma – cerebrospinal fl uid – urine

The authors declare they have no potential conflicts of interest concerning drugs, products, or services used in the study.

The Editorial Board declares that the manuscript met the ICMJE “uniform requirements” for biomedical papers.

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