Shayan P, Ebrahimzadeh E, Tageldin Mh, Amininia N, Eckert B
Iranian Research Center for Ticks and Tick-borne Diseases, Faculty of Veterinary Medicine, University of Teheran, Tehran, Iran.
Iran J Parasitol. 2011 Mar;6(1):66-72.
We used the PCR technique based on the abovementioned primer pair and sequencing to demonstrate the Theileria infection in the sheep samples collected from Sultanate of Oman.
According to the frame work of "integrated control of ticks and tick borne diseases in globalized world managed by EU-ICTTD-3 project, the samples from blood, liver, spleen, lymph node and lung were sent to the laboratory of Iranian Research Center for Ticks and Tick-borne Diseases (IRCTTD). Samples from blood smear and impression smears from liver, spleen, lymph node, and lung were analyzed by Geimsa staining. The DNA was extracted from the abovementioned samples and analyzed by PCR technique using specific primers derived from the nucleotide sequences of 18S rRNA gene of T. lestoquardi, which can amplify the common region in other Theileria and Babesia spp. Subsequently the amplified DNA was sequenced.
The analysis of blood smears of the sheep was negative for piroplasmosis performed through the Giemsa staining. The impression smears prepared from liver, spleen, lymph node, and lung showed suspicious structures mimicking Theileria schizonts in some cells. The results showed an expected PCR product of 428 bp in length, which is specific for Theileria spp. The PCR products were subsequently sequenced. The corresponding nucleotide sequence is registered under accession number JF309152 in GenBank. The sequence alignment in GenBank showed that the PCR products had 99% homology to the known T. lestoquardi registered under accession number AF081135 in the GenBank.
Oman sheep are highly susceptible for Theileria infection and the infected sheep mostly die before the microschizonts or erythrocytic form of Theileria appears in the nucleated or erytrocytic cells respectively.
我们使用基于上述引物对的PCR技术和测序方法,来证明从阿曼苏丹国采集的绵羊样本中存在泰勒虫感染。
根据欧盟ICTTD - 3项目管理的“全球化世界中蜱及蜱传疾病的综合防治”框架,将血液、肝脏、脾脏、淋巴结和肺的样本送至伊朗蜱及蜱传疾病研究中心(IRCTTD)实验室。对血液涂片以及肝脏、脾脏、淋巴结和肺的印片进行吉姆萨染色分析。从上述样本中提取DNA,并使用源自莱氏泰勒虫18S rRNA基因核苷酸序列的特异性引物,通过PCR技术进行分析,该引物可扩增其他泰勒虫属和巴贝斯虫属中的共同区域。随后对扩增的DNA进行测序。
通过吉姆萨染色对绵羊血液涂片进行梨形虫病分析,结果为阴性。从肝脏、脾脏、淋巴结和肺制备的印片中,在一些细胞中显示出类似泰勒虫裂殖体的可疑结构。结果显示出预期的长度为428 bp的PCR产物,这对泰勒虫属具有特异性。随后对PCR产物进行测序。相应的核苷酸序列已在GenBank中注册,登录号为JF309152。GenBank中的序列比对显示,PCR产物与GenBank中登录号为AF081135的已知莱氏泰勒虫具有99%的同源性。
阿曼绵羊对泰勒虫感染高度易感,且感染的绵羊大多在泰勒虫的微裂殖体或红细胞内形式分别出现在有核细胞或红细胞之前死亡。