Molecular Identification of Bloodmeal Source in Ixodes ricinus Ticks Using 12S rDNA As a Genetic Marker
Molecular Identification of Bloodmeal Source in Ixodes ricinus Ticks Using 12S rDNA As a Genetic Marker
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以 12S rDNA 作为遗传标记对蓖麻蜱血粉来源进行分子鉴定
DOI：
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发表时间：
2007
影响因子：
2.1
通讯作者：
L. Gern
中科院分区：
文献类型：
--
作者：
P. Humair;Veronique Douet;Francisca Morán Cadenas;L. Schouls;I. van de Pol;L. Gern
摘要我们建立了一种鉴定蓖麻硬蜱血粉来源的分子生物学方法，传播莱姆疏螺旋体病和蜱传脑炎的欧洲媒介。利用脊椎动物线粒体12SrDNA中约145 bp的正向延伸片段作为分子标记，对宿主脊椎动物进行了种间鉴别。该方法包括一个单一的运行聚合酶链反应扩增的12S rDNA分子标记，通过使用非简并引物，然后通过使用特定的寡核苷酸探针的反向线印迹杂交测定。探针的调色板使我们能够区分宿主脊椎动物的主要群体（例如，哺乳动物、小型啮齿动物、偶蹄类动物、鸟类、蜥蜴），并在属或种水平上鉴定血粉来源。设计外部引物并用于测序广泛范围的已知或潜在宿主脊椎动物物种（n = 60）的12S rDNA分子标记，包括哺乳动物（n = 28）、鸟类（n = 31）和爬行动物（n = 1）物种。该技术与已知的用于鉴定蜱传病原体的方法结合使用（例如，广义伯氏疏螺旋体）使我们能够确定感染性血粉的来源，并确定水库物种。本方法已成功地应用于所有饲养中血粉来源的鉴定。蓖麻蜱和半野外采集的I.蓖麻蜱。此外，在65%感染B的蜱中鉴定出血粉来源。广义上的伯多禄可以设想进一步发展这项技术，用于检测其他病媒传播的病原体及其储存宿主。
Abstract We developed an efficient molecular method for the identification of the bloodmeal sources in the tick Ixodes ricinus (L.), the European vector of the agents of Lyme borreliosis and tick-borne encephalitis. A ≈145-bp orthologous fragment of the vertebrate mitochondrial 12S rDNA was used as a molecular marker to discriminate host vertebrate species. The method consists of a single run polymerase chain reaction amplification of the 12S rDNA molecular marker by using nondegenerate primers followed by a reverse line blot hybridization assay by using specific oligonucleotide probes. The palette of probes allowed us to distinguish major groups of host vertebrates (e.g., mammals, small rodents, artiodactyls, birds, lizards) and to identify the bloodmeal sources at the genus or species level. External primers were designed and used to sequence the 12S rDNA molecular marker of a broad range of known or potential host vertebrate species (n = 60), including mammal (n = 28), bird (n = 31), and reptile (n = 1) species. The use of this technique coupled with known methods for identification of tick-borne pathogens (e.g., Borrelia burgdorferi sensu lato) allowed us to determine the source of infective bloodmeal and to identify reservoir species. The present method was successfully used to identify the source of bloodmeals in all feeding I. ricinus ticks and in half of questing field-collected I. ricinus ticks. Moreover, the bloodmeal source was identified in 65% of ticks infected with B. burgdorferi sensu lato. Further development of this technique may be envisaged for the detection of other vector-borne pathogens and their reservoir hosts.
DOI：
10.4269/ajtmh.2005.73.336
发表时间：
2005-08-01
影响因子：
3.3
作者：
Kent, RJ;Norris, DE
通讯作者：
Norris, DE
欧洲动物作为莱姆病螺旋体储存宿主的能力。
DOI：
10.1093/infdis/165.3.479
发表时间：
1992
期刊：
The Journal of infectious diseases
影响因子：
--
作者：
Matuschka,FR;Fischer,P;Heiler,M;Richter,D;Spielman,A
通讯作者：
Spielman,A
博物馆蜱虫标本肠道中哺乳动物 DNA 的检测和鉴定。
影响因子：
2.1
作者：
Tobolewski,J;Kaliszewski,MJ;Colwell,RK;OliverJr,JH
通讯作者：
OliverJr,JH