Pieniazek Norman, Sawczuk Marek, Skotarczak Bogumiła
Department of Genetics, Szczecin University, Al. Piastow 40B, 71-065 Szczecin, Poland.
J Parasitol. 2006 Feb;92(1):32-5. doi: 10.1645/GE-541R2.1.
In the present study, PCR has been applied to detect and analyze DNA of Babesia spp. extracted from Ixodes ricinus ticks. Collection of I. ricinus was made in 6 forested areas of Zachodniopomorskie Voivodship, Poland, during 2 seasonal peaks of tick activity, i.e., spring and autumn, 2001. In total, 1,328 I. ricinus were collected and processed for PCR with F34 and R323 primers. Babesia spp. was detected in 28 (2% of 1,328 tested) ticks; 26 were identified as B. divergens. The other 2 were identified as B. microti. PCR was conducted with 18S rRNA specific primers and sequencing was processed to precisely identify and compare these isolates with B. microti and B. divergens sequences from Europe, North America, and Asia obtained from the GenBank. Analysis revealed that sequences of B. microti from northwestern Poland are almost identical (99.94%) with those referred to as "Munich strain"; both form a clade different from other European strains, as well as those from Asia and North America (called B. microti, sensu stricto). An investigation performed with B. divergens sequences showed that the sequence from northwestern Poland is 99.94% homologous to an isolate from Ireland ("Purnel"), and differs in just a few nucleotides from other European sequences. Phylogenetic analysis revealed that the sequence of B divergens isolated from Polish ticks form a group that comprise 4 European sequences from Great Britain and Ireland and is, therefore, closely related to other European and North American B. divergnens sequences.
在本研究中，聚合酶链反应（PCR）已被用于检测和分析从蓖麻硬蜱中提取的巴贝斯虫属的DNA。2001年，在波兰西滨海省的6个林区，于蜱活动的两个季节高峰（即春季和秋季）采集了蓖麻硬蜱。总共采集了1328只蓖麻硬蜱，并使用F34和R323引物进行PCR检测。在28只（占检测的1328只的2%）蜱中检测到了巴贝斯虫属；其中26只被鉴定为分歧巴贝斯虫。另外2只被鉴定为微小巴贝斯虫。使用18S rRNA特异性引物进行PCR，并进行测序以精确鉴定这些分离株，并与从GenBank获得的来自欧洲、北美和亚洲的微小巴贝斯虫和分歧巴贝斯虫序列进行比较。分析表明，波兰西北部的微小巴贝斯虫序列与被称为“慕尼黑菌株”的序列几乎相同（99.94%）；两者形成一个不同于其他欧洲菌株以及来自亚洲和北美的菌株（狭义微小巴贝斯虫）的进化枝。对分歧巴贝斯虫序列进行的一项研究表明，波兰西北部的序列与来自爱尔兰的一个分离株（“珀内尔”）的同源性为99.94%，与其他欧洲序列仅在少数几个核苷酸上有所不同。系统发育分析表明，从波兰蜱中分离出的分歧巴贝斯虫序列形成一个包含来自英国和爱尔兰的4个欧洲序列的组，因此与其他欧洲和北美的分歧巴贝斯虫序列密切相关。
