title: Evaluation of a genotyping method based on the ospA gene to detect Borrelia burgdorferi sensu lato in multiple samples of lyme borreliosis patients.
abstract: In this study we have developed a new Restriction-Fragment-Length-Polymorphism (RFLP) genotyping method for rapid detection and identification of Borrelia genospecies present as unique species or as co-infection in multiple specimens obtained simultaneously from 29 individual patients affected by early or late Lyme borreliosis (LB). The target of the RFLP-genotyping was the heterogeneous plasmid located ospA gene, thus we developed a method able to detect and differentiate between six clinically relevant Borrelia genospecies circulating in Europe, B. burgdorferi sensu stricto, B. garinii, B. afzelii, B. valaisiana, B. bissettii and B. spielmanii. In this study Borrelia DNA could be detected by PCR in at least one specimen of each patient, except in one case of neuroborreliosis (NB); blood samples gave the highest sensitivity in all patient groups. The genotyping indicated that B. afzelii was present in 8 patients with skin involvement, B. garinii in 2 cases of NB and 4 cases with skin involvement, B. burgdorferi sensu stricto was detected in one patient with skin involvement and another with Lyme arthritis. Different Borrelia species in distinct specimens were identified in one patient with EM. The RFLP analysis of 11 patients revealed mixed patterns, which suggested pluri-infection with different Borrelia species.
authors: Spirochete Laboratory, Dipartimento di Scienze Biomediche, Università di Trieste, Italy.
authors: Romina
authors: R
authors: Floris
authors: Giulia
authors: G
authors: Menardi
authors: Raffaela
authors: R
authors: Bressan
authors: Giusto
authors: G
authors: Trevisan
authors: Simona
authors: S
authors: Ortenzio
authors: Emanuela
authors: E
authors: Rorai
authors: Marina
authors: M
authors: Cinco
mesh_terms: D000954:Antigens, Surface / Q000235:genetics*; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics*; D001428:Bacterial Vaccines / Q000235:genetics*; D001706:Biopsy; D025065:Borrelia burgdorferi / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D004269:DNA, Bacterial / Q000235:genetics; D003937:Diagnosis, Differential; D015787:Erythema Chronicum Migrans / Q000382:microbiology / Q000473:pathology; D005260:Female; D006801:Humans; D008074:Lipoproteins / Q000235:genetics*; D008193:Lyme Disease / Q000175:diagnosis / Q000382:microbiology* / Q000473:pathology; D020852:Lyme Neuroborreliosis / Q000382:microbiology / Q000473:pathology; D008297:Male; D008875:Middle Aged; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D012867:Skin / Q000382:microbiology / Q000473:pathology
chemical_list: D000954:Antigens, Surface; D001425:Bacterial Outer Membrane Proteins; D001428:Bacterial Vaccines; D004269:DNA, Bacterial; D008074:Lipoproteins; C072045:OspA protein