Abstract: reliable, rapid, unexpensive and specific Lyme testing

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Folia Microbiol (Praha). 2005;50(1):31-9.

Improved method of detection and molecular typing of Borrelia

burgdorferi sensu lato in clinical samples by polymerase chain

reaction without DNA purification.

Rudenko N, Golovchenko M, Nemec J, Volkaert J, Mallatova N,

Grubhoffer L.

Faculty of Biological Sciences, University of South Bohemia, 370 05

Ceske Budejovice, Czechia. natasha@...

A simple assay by polymerase chain reaction was used for the of

detection of Borrelia burgdorferi, causative agent of Lyme

borreliosis (LB).

It involves no DNA purification and is based on the amplification of

a specific region of ospA gene of B. burgdorferi, followed by direct

detection of the PCR product with SYBR Green I by agarose gel

electrophoresis.

The method was used to analyze samples from patients with LB

diagnosis, with presumable infection with the LB spirochete, those

with unclear clinical symptoms and after the course of an antibiotic

treatment.

Spirochetal DNA was detected by PCR even in contaminated samples in

which B. burgdorferi was overgrown by fungi and other bacteria.

Spirochetal DNA was detected and borrelia species was identified in

cerebrospinal fluid of two patients hospitalized with the

diagnosis " fever of unknown origin " . Western blot and ELISA were

negative in both cases.

Total analysis of 94 samples from the hospital in Ceske Budejovice

(South Bohemia, Czechia) showed infection with B. burgdorferi sensu

stricto in 11% and B. garinii in 15% of cases. The highest

prevalence was found for B. afzelii (43%). Co-infection was

confirmed in 24 % of the analyzed symplex; 7% of samples that were

B. burgdorferi sensu lato positive gave no results in DNA

amplification with B. burgdorferi sensu stricto-, B. garinii- and B.

afzelii-specific primers. The proposed reliable, rapid, unexpensive

and specific technique could form the basis of laboratory tests for

routine detection and identification of Lyme-disease spirochete in

different samples.

PMID: 15954531 [PubMed - in process]