一例并殖吸虫病患者的虫卵DNA序列分析鉴定(英文)

GENE SEQUENCING FOR IDENTIFICATION OF PARAGONIMUS EGGS FROM A HUMAN CASE

[目的 ]运用分子生物学技术分析虫卵基因序列鉴定并殖吸虫病类型。 [方法 ]先从并殖吸虫病患者痰中分离出虫卵 ,然后PCR扩增出虫卵中完整的核糖体DNA第二间隔区基因 (ITS2 ) ,并直接用于测序从而获得该基因的核苷酸序列。同时 ,亦用同法分别从动物宿主粪便中分离出的卫氏并殖吸虫和斯氏狸殖吸虫虫卵中获得ITS2基因序列作为DNA参照分析。此外 ,本文也对从患者痰中分离出的虫卵进行详细的形态学特征描述分析。 [结果 ]来自患者的虫卵ITS2基因序列与参照的卫氏并殖吸虫虫卵的基因序列 10 0 %一致 ,而与来自斯氏狸殖吸虫虫卵的基因序列只有 92 %核苷酸相同。此外 ,从形态学上讲 ,来自患者的虫卵形态特征更与卫氏并殖吸虫虫卵相似。 [结论 ]通过基因序列分析 ,可确诊患者所患的是卫氏并殖吸虫病。

Objective] To identifiy the etiologic agent from a paragonimiasis patient using molecular techniques. [Methosd] The complete nuclear ribosomal DNA second internal transcribed spacer (ITS2) gene sequence of eggs in sputum from a paragonimiasis patient was obtained by directly auto-sequencing its PCR product . ITS2 genes from eggs of Paragonimus westermani and Pagumogonimus skrjabini (both from animal hosts) were also sequenced for comparison. In addition , morphological comparisons were made with the eggs of the two species. [Results] The ITS2 gene from the human case was 100% identical with the sequence from the eggs of P westermani from an experimentally infected dog but only 92% identical with the sequence from the eggs of P skrjabini. Morphologically, the eggs from the human case more resembled those from P westermani infected dog. [Conclunsion] The patient was diagnosd to be suffered from paragonimiasis westermani by gene sequence analysis.