PCR-based methods for identification of potentially zoonotic ascaridoid parasites of the dog, fox and cat
PCR-based methods for identification of potentially zoonotic ascaridoid parasites of the dog, fox and cat
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DOI：
10.1016/s0001-706x(97)00093-4
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发表时间：
1997-11-01
期刊：
影响因子：
2.7
通讯作者：
Chilton, NB
中科院分区：
文献类型：
--
作者：
Jacobs, DE;Zhu, XQ;Chilton, NB
从狗、狐狸和猫身上收集的蛔虫线虫中提取基因组 DNA。通过 PCR 扩增每个样品核糖体 DNA 的第二个内部转录间隔区 (ITS-2) 的区域。对每种线虫物种（犬弓蛔虫、猫弓蛔虫和狮子弓蛔虫）的代表性 ITS-2 产品进行了测序。限制性位点经鉴定可用作 PCR 联锁 RFLP 测定中的遗传标记。通过使用两种核酸内切酶 HinfI 和 RsaI，可以将这三个物种彼此区分，并与人体组织中可能发现的其他蛔虫进行区分。引物针对三个物种的 ITS-2 序列的独特区域设计，用于诊断性 PCR 程序，并针对同源和异源 DNA 样本组进行评估。结果表明，这两种方法都是进一步开发用于检测和/或鉴定人体组织中蛔虫幼虫的良好候选方法。 (C) 1997 Elsevier Science B.V.
Genomic DNA was extracted from ascaridoid nematodes collected from dogs, foxes and cats. A region spanning the second internal transcribed spacer (ITS-2) of the ribosomal DNA of each sample was amplified by PCR. Representative ITS-2 products for each nematode species (Toxocara canis, Toxocara cati and Toxascaris leonina) were sequenced. Restriction sites were identified for use as genetic markers in a PCR-linked RFLP assay. The three species could be differentiated from each other and from other ascaridoids that may be found in human tissues by use of two endonucleases, HinfI and RsaI. Primers were designed to unique regions of the ITS-2 sequences of the three species for use in diagnostic PCR procedures and primer sets evaluated against panels of homologous and heterologous DNA samples. Results suggest that both methods are good candidates for further development for the detection and/or identification of ascaridoid larvae in human tissues. (C) 1997 Elsevier Science B.V.