Dunaj Justyna, Zajkowska Joanna Maria, Kondrusik Maciej, Gern Lise, Rais Oliver, Moniuszko Anna, Pancewicz Sławomir, Świerzbińska Renata
Department of Infectious Diseases and Neuroinfections, Medical University, Białystok, Poland.
nstitute of Biology, Laboratory of Eco-Epidemiology of Parasites, University of Neuchâtel, Neuchâtel, Switzerland.
Ann Agric Environ Med. 2014;21(2):239-43. doi: 10.5604/1232-1966.1108583.
RLB (Reverse Line Blot Hybridization) is a molecular biology technique that might be used for Borrelia burgdorferi sensu lato (sl) DNA detection with genospecies specification. Among B. burgdorferi sl genospecies at least 7 are regarded as pathogenic in Europe.
The aim of the study was to evaluate the frequency of different Borrelia genospecies DNA detection in Ixodes ricinus ticks in the endemic area of North-Eastern Poland by using RLB.
Ixodes ricinus ticks were collected in May - June, from 6 different sites in North-Eastern Poland (Jakubin, Kolno, Grajewo, Suwałki, Siemiatycze, Białowieża) by flagging. Extracted DNA was amplified by polymerase chain reaction (PCR) targeting the intergenic spacer 5S 23S of B. burgdorferi sl. PCR products were hybridised to 15 different oligonucleotide probes for 9 different Borrelia genospecies (B. burgdorferi sl, B. burgdorferi ss, B. garinii, B. afzelii, B. valaisiana, B. lusitaniae, B. spielmanii, B. bissettii and B. relapsing fever-like spirochetes (B. myamotoi)) by RLB.
Borrelia genospecies DNA was detected in 205 Ixodes ricinus ticks. Among 14 infected with Borrelia ticks, 4 were identified as B. garinii and 10 as B. afzelii. Higher numbers of infected ticks were noticed in the eastern part of the research area, where large forest complexes dominate. Nymphs appeared to be the most frequently infected tick stage, which has an epidemiological meaning in the incidence of Lyme borreliosis.
The study demonstrated that RLB might be easily used in Borrelia DNA detection with genospecies-identification, and indicated the domination of B. afzelii and B. garinii in ticks from North-Eastern Poland.
反向线印迹杂交（RLB）是一种分子生物学技术，可用于检测伯氏疏螺旋体狭义种（sensu lato，sl）的DNA并鉴定其基因种。在欧洲，伯氏疏螺旋体狭义种的至少7个基因种被认为具有致病性。
本研究旨在通过RLB评估波兰东北部流行地区蓖麻硬蜱中不同伯氏疏螺旋体基因种DNA的检测频率。
于5月至6月，通过拖旗法在波兰东北部的6个不同地点（雅库宾、科尔诺、格拉耶沃、苏瓦乌基、谢米亚季切、比亚沃维耶扎）采集蓖麻硬蜱。提取的DNA通过针对伯氏疏螺旋体狭义种5S-23S基因间隔区的聚合酶链反应（PCR）进行扩增。PCR产物通过RLB与针对9种不同伯氏疏螺旋体基因种（伯氏疏螺旋体狭义种、伯氏疏螺旋体指名亚种、伽氏疏螺旋体、阿氏疏螺旋体、瓦莱疏螺旋体、卢西塔尼亚疏螺旋体、斯皮尔曼疏螺旋体、比氏疏螺旋体和复发性发热样螺旋体（米亚莫托伊疏螺旋体））的15种不同寡核苷酸探针杂交。
在205只蓖麻硬蜱中检测到伯氏疏螺旋体基因种DNA。在14只感染伯氏疏螺旋体的蜱中，4只为伽氏疏螺旋体，10只为阿氏疏螺旋体。在研究区域东部发现了更多受感染的蜱，该地区以大片森林为主。若虫似乎是最常被感染的蜱阶段，这在莱姆病的发病率方面具有流行病学意义。
该研究表明RLB可轻松用于伯氏疏螺旋体DNA的检测及基因种鉴定，并表明在波兰东北部的蜱中阿氏疏螺旋体和伽氏疏螺旋体占主导地位。
