[Detection of borrelia DNA from patients with neuroborreliosis and erythema migrans].
OBJECTIVE
Assessment of PCR procedure for proving of the Borrelia burgdorferi sensu lato DNA in nerve and skin forms of Lyme borreliosis.


METHODS
DNA from plasma, urine and CSF was isolated by QIAamp DNA mini kit. PCR was designed as two-step amplification (nested-PCR). Each sample was tested in PCR for five target sequences: two were specific for plasmide genes encoding OspA and OspC proteins and three correlated with genes for 16SrDNA, flagellin and p66 protein.


RESULTS
Borrelial DNA was proved in 41 patients suffering from neuroborreliosis out of 56 (77.4 %), among 48 patients with erythema migrans (EM) were found 26 positive (54.2 %). After treatment the specific DNA was detected in 22 patients with neuroborreliosis (41.5 %) and 16 patients with EM (38.1 %). Three months after the treatment 23 patients were positive in both of groups (28.7 %) and next 3 months later the specific DNA was found in 6 (9.5 %). The highest rate of positive results was manifested by 16SrDNA target, lower and comparable results were obtained by OspA, C and flagellin primers, the lowest rate was in p66 system.


CONCLUSIONS
The tested PCR proved specific DNA in all tested biological fluids in both of the clinical forms of Lyme borreliosis with a relatively high sensitivity. The proving of DNA can not be used for the assessment of the effect of treatment due to the long persistence of PCR positivity after antibiotic treatment. To achieve a sufficient diagnostic sensitivity of PCR it is desirable to use minimally two amplification systems in parallel.
Moravcová
Lenka
L
Department of Infectious Diseases, Charles University Prague, Czech Republic. Lenka.moravcova@fnb.cz
Pícha
Dusan
D
Vanousová
Daniela
D
Hercogová
Jana
J
D015748:Borrelia burgdorferi Group / Q000235:genetics / Q000302:isolation & purification*; D004269:DNA, Bacterial / Q000032:analysis*; D015787:Erythema Chronicum Migrans / Q000175:diagnosis*; D006801:Humans; D020852:Lyme Neuroborreliosis
D004269:DNA, Bacterial