利用两对检测16S rRNA和ankA基因的引物，通过聚合酶链反应测定蓖麻硬蜱中嗜吞噬细胞无形体的患病率。

Prevalence of Anaplasma phagocytophilum in Ixodes ricinus ticks determined by polymerase chain reaction with two pairs of primers detecting 16S rRNA and ankA genes.

作者信息

Chmielewska-Badora Jolanta, Zwoliński Jacek, Cisak Ewa, Wójcik-Fatla Angelina, Buczek Alicja, Dutkiewicz Jacek

机构信息

Department of Occupational Biohazards, Institute of Agricultural Medicine, Jaczewskiego 2, 20-090 Lublin, Poland.

出版信息

Ann Agric Environ Med. 2007;14(2):281-5.

PMID:18247465

Abstract

A total of 684 Ixodes ricinus ticks (321 nymphs, 184 males, and 179 females) were collected by flagging lower vegetation in 6 forest districts located on the territory of Lublin province (eastern Poland). Ticks were examined by polymerase chain reaction (PCR) method for the presence of Anaplasma phagocytophilum DNA with two pairs of primers: EHR521/EHR747 for detecting 16S rRNA gene, and LA6/LA1 for detecting ankA gene. To study the relationship between infection in ticks and people occupationally exposed to tick bite, blood serum samples of 261 forestry workers employed in the same forest districts were examined by immunofluorescence method for the presence of specific antibodies against A. phagocytophilum. A total of 70 ticks out of 684 examined (10.2%) showed the presence of A. phagocytophilum 16S rRNA gene. The prevalence of infection was significantly dependent on tick's stage (chi-square=49.2, p<0.00001) and geographical locality (chi-square=34.4, p<0.00001). The percentage of I. ricinus females infected with A. phagocytophilum (24.6%) was significantly greater compared to males (6.5%) and nymphs (4.4%) (p<0.00001). Only 19 ticks out of 684 examined (2.8%) showed the presence of A. phagocytophilum ankA gene, significantly less compared to 16S rRNA gene (p<0.00001). The prevalence of infection demonstrated by the presence of ankA gene was also significantly dependent on tick's stage (chi-square=23.6, p<0.00001) but not on locality (chi-square=9.8, p=0.082). A significant correlation was found between the presence of A. phagocytophilum 16S rRNA gene in I. ricinus female ticks from the particular forest districts and the serologic response to A. phagocytophilum of forestry workers employed in the same districts (p<0.05). No significant correlation was found between the presence of A. phagocytophilum ankA gene in I. ricinus ticks and serologic response of exposed workers. In conclusion, detection of A. phagocytophilum infection in ticks by PCR with the use of EHR521/EHR747 primers detecting 16S rRNA gene is significantly more sensitive compared to LA6/LA1 primers detecting ankA gene.

摘要

通过在波兰东部卢布林省境内的6个森林区，对低矮植被进行拖旗法采集，共收集到684只蓖麻硬蜱（321只若虫、184只雄蜱和179只雌蜱）。采用聚合酶链反应（PCR）方法，使用两对引物检测蜱中嗜吞噬细胞无形体DNA：EHR521/EHR747用于检测16S rRNA基因，LA6/LA1用于检测ankA基因。为研究蜱感染与职业性接触蜱叮咬人群之间的关系，采用免疫荧光法检测了同一森林区261名林业工人的血清样本中抗嗜吞噬细胞无形体特异性抗体的存在情况。在684只被检测的蜱中，共有70只（10.2%）显示存在嗜吞噬细胞无形体16S rRNA基因。感染率显著取决于蜱的阶段（卡方=49.2，p<0.00001）和地理位置（卡方=34.4，p<0.00001）。感染嗜吞噬细胞无形体的蓖麻硬蜱雌蜱百分比（24.6%）显著高于雄蜱（6.5%）和若虫（4.4%）（p<0.00001）。在684只被检测的蜱中，只有19只（2.8%）显示存在嗜吞噬细胞无形体ankA基因，与16S rRNA基因相比显著较少（p<0.00001）。由ankA基因存在所显示的感染率也显著取决于蜱阶段（卡方=23.6，p<0.00001），但不取决于地理位置（卡方=9.8，p=0.082）。在特定森林区的蓖麻硬蜱雌蜱中嗜吞噬细胞无形体16S rRNA基因的存在与同一地区林业工人对嗜吞噬细胞无形体的血清学反应之间发现显著相关性（p<0.05）。在蓖麻硬蜱中嗜吞噬细胞无形体ankA基因的存在与暴露工人的血清学反应之间未发现显著相关性。总之，与检测ankA基因的LA6/LA1引物相比，使用检测16S rRNA基因的EHR521/EHR747引物通过PCR检测蜱中嗜吞噬细胞无形体感染显著更敏感。