Isolation of Borrelia burgdorferi from ticks in the Highlands of Scotland

Identification of different Borrelia burgdorferi genomic groups from Scottish ticks

“…An in house ELISA is also available using antigen from the same reference strain as is used for immunoblotting. 10 Although B burgdorferi sensu stricto was found in our study, the RAPD patterns were not identical to those of the European strain used in our in house ELISA and immunoblotting techniques. Therefore, local strains of B burgdorferi sensu stricto and B afzelii could improve the sensitivity and specificity of our in house serological tests.…”

“…8 9 Recently, 12 B burgdorferi sensu lato isolates were successfully cultured from ticks collected in one location in the Highlands of Scotland. 10 Our study set out to characterise the 12 Highland isolates using three molecular methods: an outer surface membrane protein A (OSP A) gene polymerase chain reaction (PCR) designed to give diVerent molecular weight products with diVerent genomic groups, randomly amplified polymorphic DNA (RAPD) analysis, and a PCR method using genomic group specific primers for the ribosomal RNA (rRNA) gene. [11][12][13] Methods CULTURE AND PREPARATION OF BORRELIA Twelve isolates of B burgdorferi were cultured from ticks collected on three separate occasions in the summer of 1997 from two locations approximately five and 10 miles to the east of Inverness in the Highlands of Scotland.…”

“…[11][12][13] Methods CULTURE AND PREPARATION OF BORRELIA Twelve isolates of B burgdorferi were cultured from ticks collected on three separate occasions in the summer of 1997 from two locations approximately five and 10 miles to the east of Inverness in the Highlands of Scotland. 10 Both locations were wooded and the ticks were collected by sheet dragging the grassy areas under the trees. A total of 128 ticks were collected, giving an overall positivity rate of 9.4%.…”

“…Cultures were incubated at 33°C under microaerophilic conditions. 10 The organisms were harvested by centrifugation at 10 000 ×g for 30 minutes and the pellets washed twice in 0.9% (wt/vol) sodium chloride (Steripak, Runcorn, Cheshire, UK). They were then centrifuged again at 10 000 ×g for 30 minutes and resuspended in minimal amounts of sterile distilled water.…”

Morphoea and Borrelia burgdorferi: results from the Scottish Highlands in the context of the world literature

“…12 The first stage PCR was performed with 20 µl of sample in a reaction volume of 50 µl, containing final concentrations of 10mM Tris/HCl (pH 8.5), 50mM KCl 2 , 3.5mM MgCl 2 (Applied Biotechnologies), 0.2mM dNTP (Pharmacia Biosystems, Milton Keynes, UK), 0.5 U Taq polymerase (Applied Biotechnologies), and 0.5 µM of each primer F1 (5′-ATT AAC GCT GCT AAT CTT AGT-3′) and F3 (5′-GTA CTA TTC TTT ATA GAT TC-3′) (Severn Biotech, Kidderminster, UK). The thermal cycling conditions were 35 cycles of one minute at 94°C, two minutes at 41°C, and three minutes at 66°C, followed by a further extension period of five minutes at 72°C.…”