Application of medical and analytical methods in Lyme borreliosis monitoring

Abstract

Lyme borreliosis (LB) is one of the most common tick-borne diseases in the northern hemisphere. It is a chronic inflammatory disease caused by the spirochaete Borrelia burgdorferi. In its early stages, pathological skin lesions, namely erythema chronicum migrans, appear. The lesions, usually localised at the site of the bite, may become visible from a few weeks up to 3 months after the infection. Predominant clinical symptoms of the disease also involve joint malfunctions and neurological or cardiac disorders. Lyme disease, in all its stages, may be successfully treated with antibiotics. The best results, however, are obtained in its early stages. In order to diagnose the disease, numerous medical or laboratory techniques have been developed. They are applied to confirm the presence of intact spirochaetes or spirochaete components such as DNA or proteins in tick vectors, reservoir hosts or patients. The methods used for the determination of LB biomarkers have also been reviewed. These biomarkers are formed during the lipid peroxidation process. The formation of peroxidation products generated by human organisms is directly associated with oxidative stress. Apart from aldehydes (malondialdehyde and 4-hydroxy-2-nonenal), many other unsaturated components such as isoprostenes and neuroprostane are obtained. The fast determination of these compounds in encephalic fluid, urine or plasma, especially in early stages of the disease, enables its treatment. Various analytical techniques which allow the determination of the aforementioned biomarkers have been reported. These include spectrophotometry as well as liquid and gas chromatography. The analytical procedure also requires the application of a derivatization step by the use of selected reagents.

Fig. 1

Formation of MDA during peroxidation of PUFAs [25]

Fig. 2

Pathways of PUFA peroxidation and the formation of HNE [26]

Fig. 3

Proposed pathways of lipid peroxidation which lead to the synthesis of isoprostanes [24]

Fig. 4

Formation of various classes of compounds from AA [34]

Fig. 5

Formation of MDA–TBA₂ adduct after reaction between MDA and TBA [25]

Fig. 6

Content of HNE in venous blood stored in ice vs. time of storage [87]. Methodology: after 3 h of derivatization with DNPH the concentration of HNE was determined by HPLC