Li M W, Lin R Q, Chen H H, Sani R A, Song H Q, Zhu X Q
Laboratory of Parasitology, College of Veterinary Medicine, South China Agricultural University, 483 Wushan Street, Tianhe District, Guangzhou 510642, Guangdong Province, The People's Republic of China.
Mol Cell Probes. 2007 Oct-Dec;21(5-6):349-54. doi: 10.1016/j.mcp.2007.04.004. Epub 2007 Apr 22.
Based on the sequences of the internal transcribed spacers (ITS-1 and ITS-2) of nuclear ribosomal DNA (rDNA) of Toxocara canis, Toxocara cati, Toxocara malaysiensis and Toxascaris leonina, specific forward primers were designed in the ITS-1 or ITS-2 for each of the four ascaridoid species of dogs and cats. These primers were used individually together with a conserved primer in the large subunit of rDNA to amplify partial ITS-1 and/or ITS-2 of rDNA from 107 DNA samples from ascaridoids from dogs and cats in China, Australia, Malaysia, England and the Netherlands. This approach allowed their specific identification, with no amplicons being amplified from heterogeneous DNA samples, and sequencing confirmed the identity of the sequences amplified. The minimum amounts of DNA detectable using the PCR assays were 0.13-0.54ng. These PCR assays should provide useful tools for the diagnosis and molecular epidemiological investigations of toxocariasis in humans and animals.
基于犬弓首蛔虫、猫弓首蛔虫、马来西亚弓首蛔虫和狮弓蛔虫核糖体DNA（rDNA）的内转录间隔区（ITS-1和ITS-2）序列，针对犬猫的四种蛔科动物，在ITS-1或ITS-2中设计了特异性正向引物。这些引物分别与rDNA大亚基中的一个保守引物一起使用，以扩增来自中国、澳大利亚、马来西亚、英国和荷兰的犬猫蛔科动物的107个DNA样本中的rDNA部分ITS-1和/或ITS-ITS-2。这种方法能够对它们进行特异性鉴定，异源DNA样本未扩增出扩增子，测序证实了扩增序列的身份。使用PCR检测可检测到的DNA最小量为0.13 - 0.54纳克。这些PCR检测方法应为人类和动物弓首蛔虫病的诊断和分子流行病学调查提供有用的工具。
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