Rar V A, Epikhina T I, Livanova N N, Panov V V, Doroshenko E K, Pukhovskaia N M, Vysochina N P, Ivanov L I
Mol Gen Mikrobiol Virusol. 2011(2):17-23.
A total of 3552 Ixodes persulcatus from Sverdlovsk, Chelyabinsk, Novosibirsk, Irkutsk regions and Khabarovsk Territory were examined on the Ehrlichia and Anaplasma presence by nested PCR based on the 16S rRNA gene. Both Anaplasma phagocytophilum and Ehrlichia muris DNA were found in I. persulcatus in all studied regions. A. Phagocytophilum was detected in 1.3-6.3% of ticks and E. muris - in 2.0-14.1% of ticks. Moreover, "Candidatus Neoehrlichia mikurensis" DNA was found in 8 ticks collected in Novosibirsk, Irkutsk Regions and Khabarovsk Territory. Partial nucleotide sequences of 16S rRNA gene and groESL operone (1240-1300 bp) were determined for 65 samples of A. Phagocytophilum, 17 samples of E. muris and 4 samples of "Candidatus Neoehrlichia mikurensis". Nucleotide sequences of 16S rRNA gene and groESL operone of E. muris and "Candidatus Neoehrlichia mikurensis" were shown to be highly conservative, and nucleotide sequences of groESL operone of both E. muris and "Candidatus Neoehrlichia mikurensis" differed from the sequences found previously in other species of Ixodid tick. On the basis of analysis of the 16S rRNA gene and groESL operone sequences it was concluded that all revealed samples A. Phagocytophilum could be divided into 2 groups. GroESL operone sequences of A. Phagocytophilum from the first group were identical to each other but significantly differed from the known groESL operone sequences (less than 98.2% of similarity), whereas their 16S rRNA gene sequences were identical to the sequence of widely distributed and pathogenic for human A. Phagocytophilum genetic variant (CAHU-HGEl, GenBank AF093788) or differed from it by a single nucleotide substitution. The nucleotide sequences of groESL operone of A. Phagocytophilum from the second group differed from each other by 1-4 nucleotides and were closely related (99.2-99.4% of similarity) to the sequences of groESL operone ofA. phagocytophilum isolates found in Europe in Ixodes ricinus and roe deer. The nucleotide sequences of the 16S rRNA gene of A. Phagocytophilum from the second group were most similar to the sequence of the rare A. Phagocytophilum genetic variant previously found only in China (GenBank DQ342324).
对来自斯维尔德洛夫斯克州、车里雅宾斯克州、新西伯利亚州、伊尔库茨克州和哈巴罗夫斯克边疆区的总共3552只全沟硬蜱，基于16S rRNA基因通过巢式PCR检测埃立克体属和无形体属。在所有研究地区的全沟硬蜱中均发现了嗜吞噬细胞无形体和鼠埃立克体的DNA。检测到1.3% - 6.3%的蜱带有嗜吞噬细胞无形体，2.0% - 14.1%的蜱带有鼠埃立克体。此外，在新西伯利亚州、伊尔库茨克州和哈巴罗夫斯克边疆区采集的8只蜱中发现了“新埃立克体候选种”的DNA。对65份嗜吞噬细胞无形体样本、17份鼠埃立克体样本和4份“新埃立克体候选种”样本测定了16S rRNA基因和groESL操纵子（1240 - 1300 bp）的部分核苷酸序列。结果表明，鼠埃立克体和“新埃立克体候选种”的16S rRNA基因和groESL操纵子的核苷酸序列高度保守，且鼠埃立克体和“新埃立克体候选种”的groESL操纵子的核苷酸序列与之前在其他硬蜱属物种中发现的序列不同。基于对16S rRNA基因和groESL操纵子序列的分析得出结论，所有检测出的嗜吞噬细胞无形体样本可分为两组。第一组嗜吞噬细胞无形体的groESL操纵子序列彼此相同，但与已知的groESL操纵子序列显著不同（相似度低于98.2%），而它们的16S rRNA基因序列与广泛分布且对人类致病的嗜吞噬细胞无形体基因变体（CAHU - HGEl，GenBank AF093788）的序列相同，或仅相差一个核苷酸替换。第二组嗜吞噬细胞无形体的groESL操纵子的核苷酸序列彼此相差1 - 4个核苷酸，并且与在欧洲蓖麻硬蜱和狍中发现的嗜吞噬细胞无形体分离株的groESL操纵子序列密切相关（相似度为99.2% - 99.4%）。第二组嗜吞噬细胞无形体的16S rRNA基因的核苷酸序列与之前仅在中国发现的罕见嗜吞噬细胞无形体基因变体的序列最为相似（GenBank DQ342324）。
