Detection and Differentiation of Theileria annulata and Theileria parva Using Macroschizont‐derived DNA Probes
Detection and Differentiation of Theileria annulata and Theileria parva Using Macroschizont‐derived DNA Probes
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使用源自巨裂殖体的 DNA 探针检测和区分环状泰勒虫和细小泰勒虫
DOI：
10.1111/j.1749-6632.1998.tb11038.x
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发表时间：
1998
影响因子：
5.2
通讯作者：
J. Ahmed
中科院分区：
文献类型：
--
作者：
P. Shayan;R. Biermann;E. Schein;J. Gerdes;J. Ahmed
摘要：构建了一个基于T.用在兔中产生的抗血清对部分纯化的T. annulata菌株进行筛选。检测到两个克隆，测序并命名为SA 288和SB 288（Shayan等人，出版）。根据这两个基因的序列设计寡核苷酸引物，通过聚合酶链反应（PCR）特异性扩增寄生虫DNA。我们可以证明这些基因是寄生源的，并且确实存在于所有的T。annulata股票测试在本研究中。此外，SA 288的靶序列也可以在小锥虫中鉴定。它们均不与不同种类的巴氏杆菌基因组DNA发生反应。第三对引物是从编码小锥虫特异性酪蛋白激酶II-α亚基的基因的DNA序列设计的。使用此引物对，靶序列只能在小锥虫中检测到。综上所述，本文所述的引物可用作PCR中检测泰勒虫寄生虫和区分环形泰勒虫与小泰勒虫的分子工具。
ABSTRACT: A λgt11 expression library based on T. annulata‐infected cells was screened with an antiserum raised in rabbits against partially purified schizonts of T.annulata. Two clones were detected, sequenced and designated as SA288 and SB288 (Shayan et al., submitted for publication). From the sequences of these two genes oligonucleotide primers were designed for specific amplification of parasite DNA by polymerase chain reaction (PCR). We could show that these genes are of parasitic origin and do occur in all T. annulata stocks tested in the present study. In addition, a target sequence for SA288 could also be identified in T.parva‐schizonts. None of them reacted with genomic DNA of different Babesia spp. A third primer pair was designed from the DNA‐sequence of a gene encoding for the T.parva‐specific casein kinase II‐alpha subunit. Using this primer pair, a target sequence could only be detected in T.parva. Taken together, the primers described here can be used as molecular tools in PCR for the detection of Theileria parasites and to distinguish T.annulata from T.parva.