福建省蛇舌形虫18S rRNA基因的扩增及虫种鉴定  

Amplification of 18S rRNA Gene and Identification of Pentastomiasis from Snake in Fujian Province

作　　者：宋肇程 黄潇航 夏锡锐 黄志坚[1,2] 殷光文 SONG Zhao-cheng;HUANG Xiao-hang;XIA Xi-rui;HUANG Zhi-jian;YIN Guang-wen(College of Animal Science,Fujian Agriculture and Forestry University,Fuzhou 350002,China;Fujian Animal Medicine Engineering Laboratory,Fuzhou 350002,China;College of Veterinary Medicine,South China Agricultural University,Guangzhou 510642,China)

机构地区：[1]福建农林大学动物科学学院,福建福州350002 [2]福建省动物药物工程实验室,福建福州350002 [3]华南农业大学兽医学院,广东广州510642

出　　处：《中国兽医杂志》2020年第7期112-115,I0016,共5页Chinese Journal of Veterinary Medicine

基　　金：国家自然科学基金面上项目(31872466);福建省科技厅引导性项目(2019N0005);福建农林大学科技创新专项基金项目(KF2015017,CXZX2017235,CXZX2017487)。

摘　　要：以我国福建省三明市某蛇场的病蛇为研究对象,对其进行剖检,观察其器官组织病变,并从体腔中分离得到1对寄生虫成虫,经过剖检观察初步怀疑为舌形虫病。提取虫体和肠道结节DNA,利用18S rRNA基因特异性的引物对提取的DNA进行PCR扩增,经琼脂糖凝胶电泳得到目的条带,切胶回收纯化,连接转化培养得到单菌落。对菌落进行PCR扩增测序。结果显示:病蛇肺脏、肠道病变严重有肿大、淤血和大量结节。经测序得到序列总长均为770 bp,与NCBI公布的相关舌形虫18S rRNA基因序列进行比对分析,三明蛇场得到的舌形虫成虫和肠道结节的DNA序列与EU370434.1(Raillietiella sp.1 BMR-2008 strain C129);AY744887.1(Raillietiella sp.MCZ DNA101107)的相似性为99.74%;与KC904945.1(Raillietiella orientalis)的相似性为97.40%;与JX088397.2(Linguatula serrate isolate NoDog1)的相似性为93.51%。表明此次分离的舟山眼镜蛇舌形虫属于Raillietiella属舌形虫。A diseased snake from a snake farm in Sanming city,Fujian province,was dissected to observe the lesions of its organs and tissues.Then,a pair of parasites were discovered from the body cavity of the snake.DNA was extracted from the worms and the intestinal nodule.The extracted DNA was amplified by PCR using 18S rRNA gene specific primers,and the target band was obtained by agarose gel electrophoresis.DNA was recovered and purified,and was then ligated to the carrier and transformed into the competent cells.The competent cells were cultured to obtain a single colony.The colony was amplified and sequenced.The results showed that the pulmonary and intestinal lesions of the diseased snake were characterized by swelling,congestion and a large number of nodules.The sequencing results showed that the total length of 18S rRNA gene sequences of the worms and the nodule were 770 bp.The sequences were compared with the 18S rRNA gene sequence published in NCBI.The similarity to EU370434.1(Raillietiella sp.1 BMR-2008 strain C129)and AY744887.1(Raillietiella sp.MCZ DNA101107)was 99.74%.The similarity to KC904945.1(Raillietiella orientalis)was 97.40%.The similarity to JX088397.2(Linguatula serrate isolate NoDog1)was 93.51%.The results of the study prove that the worms isolated from Naja atra is the pentastomid of the Raillietiella.

关 键 词：舟山眼镜蛇 舌形虫 18S rRNA基因序列 人兽共患病 

分 类 号：R285.5[医药卫生—中药学]