云南省血吸虫病流行区一种拟钉螺及其寄生吸虫尾蚴的分子鉴定  

Molecular identification of Tricula spp.and the parasitized trematode cercariae in schistosomiasis-endemic areas of Yunnan Province

作　　者：杜春红[1] 吕山[2] 张云[1] 李石柱[2] 熊孟韬[1] 何志海 李志华 吴明寿[1] 孙佳昱[1] 任银本 陈春琼 顾琼 王云松 董毅[1] DU Chun⁃Hong;LÜ Shan;ZHANG Yun;LI Shi⁃Zhu;XIONG Meng⁃Tao;HE Zhi⁃Hai;LI Zhi⁃Hua;WU Ming⁃Shou;SUN Jia⁃Yu;REN Yin⁃Ben;CHEN Chun⁃Qiong;GU Qiong;WANG Yun⁃Song;DONG Yi(Yunnan Institute of Endemic Diseases Control and Prevention,Dali 671000,China;National Institute of Parasitic Diseases,Chinese Center for Disease Control and Prevention,Chinese Center for Tropical Diseases Research,China;Xiangyun Station of Schistosomiasis Control,Yunnan Province,China)

机构地区：[1]云南省地方病防治所,大理671000 [2]中国疾病预防控制中心寄生虫病预防控制所、国家热带病研究中心 [3]云南省祥云县血吸虫病防治站

出　　处：《中国血吸虫病防治杂志》2020年第2期159-167,共9页Chinese Journal of Schistosomiasis Control

摘　　要：目的对云南省血吸虫病流行区的一种拟钉螺及其寄生吸虫进行分子生物学鉴定,了解其种属分类地位。方法采集云南省祥云县拟钉螺和钉螺,压碎镜检采集螺软体腹足部组织,并观察其体内尾拗寄生情况。采用逸拗法采集拟钉螺寄生的不同形态尾拗,螺体组织和尾拗样本分别提取基因组DNA,采用PCR法扩增螺体内16S核糖体RNA(16S rRNA)、细胞色素c氧化酶I(COI)、28S核糖体DNA(28S rDNA)基因及其体内寄生吸虫尾拗的NADH脱氢酶亚基1(ND1)、28S rfNA基因,对扩增产物进行测序,并通过序列比对及系统进化分析鉴定样本螺和寄生吸虫种属。结果共检测拟钉螺样本382只,发现无叉形、双叉形、燕子形等3种形态吸虫尾拗,阳性率分别为20.94%(80/382)、3.40%(13/382)、7.07%(27/382)。基于16S rRNA、COI、28S rDNA基因的分子进化分析显示,该拟钉螺与滇池德拉维螺(Delavaya dianchiensis)同源性较高、聚在同一个分支。基于ND1、28S rDNA4基因的序列分析显示,无叉形尾拗吸虫属侧殖吸虫科,燕子形尾拗吸虫属孔肠科,双叉形尾拗吸虫可能为不同于中华血吸虫(Schistosoma sinensium)和勾形卵血吸虫(Schistosoma ovuncatum)的另一种血吸虫。结论初步了解了云南省血吸虫病流行区拟钉螺及其寄生吸虫种属分类,但更明确的分类关系和危害性等尚有待深入研究。Objective To characterize a species of the genus Tricula and parasitized trematodes in schistosomiasis-endemic areas of Yunnan Province using a molecular analysis,so as to understand their taxonomic positions.Methods Tricula spp.and Oncomelania snails were collected from Xiangyun County,Yunnan Province,and cercaria parasitizing snails were observed using crushing followed by microscopy.Cercaria parasitizing Tricula snails at various morphologies were sampled using a shedding method.Genomic DNA was extracted from snail soft tissues and cercariae,and the 16 S rRNA,COI,28 S rDNA genes in snails and the ND1 and 28 S rDNA genes in cercariae were amplified using a PCR assay and sequenced.The species of Tricula snails and their parasitized trematodes was characterized using sequence alignment and phylogenetic analysis.Results Among 382 Tricula snails detected,there were three types of trematode cercariae found,including the non-forked(20.94%,80/382),double-forked(3.40%,13/382)and swallo-w shapes(7.07%,27/382).Sequence and phylogenetic analyses showed that the 16 S rRNA,COI and28 S rDNA gene sequences of this species of Tricula had high homology to those in Delavaya dianchiensis,and were clustered in a branch.Sequencing analysis of the ND1 and 28 S rDNA genes revealed that the non-forked cercariae belonged to the family Pleurogenidae,the swallow-shaped cercariae belonged to the family Opecoelidae,and the double-forked cercariae belonged to another species of the genus Schistosoma that was different from S.sinensium and S.ovuncatum.Conclusion The species and taxonomy of Triculla spp.and their parasitized trematodes are preliminarily determined in schistosomiasis-endemic areas of Yunnan Province;however,further studies are required to investigate the more definite taxonomy and pathogenicity.

关 键 词：拟钉螺 钉螺 吸虫 尾蚴 16SRRNA基因 COI基因 28SrDNA基因 ND1基因 血吸虫病 云南省 

分 类 号：R383.24[医药卫生—医学寄生虫学]