1例蓝氏贾第鞭毛虫感染者的虫种分子鉴定及基因溯源

doi:10.12140/j.issn.1000-7423.2023.03.018

中国寄生虫学与寄生虫病杂志 ›› 2023, Vol. 41 ›› Issue (3): 380-383.doi: 10.12140/j.issn.1000-7423.2023.03.018

1例蓝氏贾第鞭毛虫感染者的虫种分子鉴定及基因溯源

王丹¹^,²(), 贺志权¹^,², 刘颖¹^,², 刘灵芝³, 陈慧慧⁴, 蒋甜甜¹^,², 纪鹏慧¹^,², 钱丹¹^,², 杨成运¹^,², 张红卫¹^,²^,^*()

1 河南省疾病预防控制中心，郑州 450016
2 河南省传染病病原生物重点实验室，郑州 450016
3 郑州大学附属儿童医院，河南省儿童医院，郑州儿童医院，郑州 450000
4 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），国家卫生健康委员会寄生虫病原与媒介生物学重点实验室，世界卫生组织热带病合作中心，国家级热带病国际联合研究中心，上海 200025

收稿日期:2022-07-21 修回日期:2023-01-23 出版日期:2023-06-30 发布日期:2023-06-15
通讯作者: *张红卫（1969-），男，博士，主任医师，从事寄生虫病防治和研究等工作。E-mail：zhwei69@163.com
作者简介:王丹（1994-），女，硕士，技师，从事寄生虫病防治和研究工作。E-mail：2859013855@qq.com
基金资助:
河南省医学科技攻关计划联合共建项目(LHGJ20220178)

Molecular identification and genetic tracing of Giardia lamblia isolated from an infected case

WANG Dan¹^,²(), HE Zhiquan¹^,², LIU Ying¹^,², LIU Lingzhi³, CHEN Huihui⁴, JIANG Tiantian¹^,², JI Penghui¹^,², QIAN Dan¹^,², YANG Chengyun¹^,², ZHANG Hongwei¹^,²^,^*()

1 Henan Centre for Disease Control and Prevention, Zhengzhou 450016, China
2 Henan Provincial Key Laboratory for Infectious Disease Prevention and Control, Zhengzhou 450016, China
3 Children’s Hospital affiliated of Zhengzhou University; Henan Children’s Hospital; Zhengzhou Children’s Hospital, Zhengzhou 450000, China
4 National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention (Chinese Center for Tropical Diseases Research); NHC Key Laboratory of Parasite and Vector Biology; WHO Collaborating Centre for Tropical Diseases; National Center for International Research on Tropical Diseases, Shanghai 200025, China

Received:2022-07-21 Revised:2023-01-23 Online:2023-06-30 Published:2023-06-15
Contact: *E-mail: zhwei69@163.com
Supported by:
Henan Medical Science and Technology Plan Project(LHGJ20220178)

摘要/Abstract

摘要：

对1例蓝氏贾第鞭毛虫感染者进行虫种分子鉴定并分析其感染来源。收集患儿与其父母、保姆的新鲜粪样，碘液染色后镜检，提取粪样DNA，采用巢式PCR扩增贾第虫磷酸丙糖异构酶（tpi）、谷氨酸脱氢酶（gdh）、β-贾第素（bg）基因并测序，通过BLAST、ChromasPro和MEGA 11.0等软件对基因序列进行系统进化分析并判断其集聚体类型。结果显示，镜下可见患儿粪样中有贾第虫滋养体和包囊。巢式PCR均扩增出约500 bp的条带，与贾第虫属tpi、gdh、bg基因片段一致，确认该患儿为贾第虫感染；患儿父母和保姆均无腹泻症状，但在其父亲粪样中发现贾第虫包囊，结合巢式PCR与测序结果分析，其父为贾第虫带虫者。基因比对分析结果显示，患儿父子二人粪样扩增出的tpi、gdh、bg基因序列一致性分别为99.8%、100%和98.5%，其中tpi基因序列与贾第虫集聚体AⅡ型（GenBank登录号：LC183963）的序列一致性分别为100%、99.8%，gdh基因序列与贾第虫集聚体AⅡ型（GenBank登录号：KF843931）的序列一致性分别为99.6%、100%，患儿bg基因序列与集聚体AⅢ型（GenBank登录号：LC183968）的序列一致性为99.2%，患儿父亲bg基因序列与集聚体AⅡ型（GenBank登录号：LC183975）的序列一致性为100%。系统进化树分析结果显示，患儿与其父亲所感染的贾第虫均与集聚体A型虫株聚在一支，且以tpi和gdh为靶基因进行亚型分析均与贾第虫集聚体AⅡ型聚在一支。由此判断患儿为贾第虫集聚体AⅡ型感染，且可能为家庭内部传播导致的家庭聚集性感染。

关键词: 蓝氏贾第鞭毛虫, 磷酸丙糖异构酶基因, 谷氨酸脱氢酶基因, β-贾第素基因, 系统进化分析

Abstract:

To investigate the molecular identification of Giardia lamblia isolated from an infected case and analyze the source of infection. Fresh fecal samples of the patient and his parents and the nanny were collected for iodine solution staining and microscopic examination. Fecal DNA was extracted, and nested PCR was used to amplify the triosephosphate isomerase (tpi), glutamate dehydrogenase (gdh) and beta-giardin (bg) gene for sequencing. The homology comparison and phylogenetic analysis of gene sequences were performed by BLAST, ChromasPro and MEGA 11.0 software to determine the assemblages. The results showed that there were trophozoites and cysts in the fecal of the patient through microscopic examination. Nested PCR amplified bands of about 500 bp, which were consistent with the tpi, gdh and bg gene fragments of G. lamblia, confirming the infection of G. lamblia. None of the patient’s parents and nanny had diarrhea symptoms, but G. lamblia cysts were found in the stool specimen of his father. Combined with nested PCR and sequencing results, the father was a G. lamblia carrier. The results of the gene comparison showed that the percent identity of tpi, gdh and bg genes sequence amplified from fecal samples of the patient and his father was 99.8%, 100% and 98.5%, respectively. The percent identity of tpi gene sequence amplified from the fecal samples of the patient and his father showed 100% and 99.8% with G. lamblia assemblages AⅡ (GenBank accession no. LC183963), respectively. The amplified gdh gene sequence showed 99.6% and 100% with assemblages AⅡ (GenBank accession no. KF843931), respectively. The percent identity between the bg gene sequence amplified from the patient fecal sample and the sequence of assemblages AⅢ (GenBank accession no. LC183968) is 99.2%, while the percent identity between the bg gene sequence amplified from the father’s fecal sample and the sequence of assemblages AⅡ (GenBank accession no. LC183975) is 100%. The results of phylogenetic tree analysis showed that both the patient and his father were infected with G. lamblia and clustered in the same cluster as assemblages A, and sub-genotype analysis using tpi and gdh as target genes were conducted, which clustered in the same cluster as assemblages AⅡ. Therefore, it can be determined that the patient was infected with G. lamblia assemblages AⅡ, and it could be a family cluster infection caused by internal transmission within the family.

Key words: Giardia lamblia, Triosephosphate isomerase genes, Glutamate dehydrogenase gene, Beta-giardin gene, Phylogenetic analysis

中图分类号:

382.213

王丹, 贺志权, 刘颖, 刘灵芝, 陈慧慧, 蒋甜甜, 纪鹏慧, 钱丹, 杨成运, 张红卫. 1例蓝氏贾第鞭毛虫感染者的虫种分子鉴定及基因溯源[J]. 中国寄生虫学与寄生虫病杂志, 2023, 41(3): 380-383.

WANG Dan, HE Zhiquan, LIU Ying, LIU Lingzhi, CHEN Huihui, JIANG Tiantian, JI Penghui, QIAN Dan, YANG Chengyun, ZHANG Hongwei. Molecular identification and genetic tracing of Giardia lamblia isolated from an infected case[J]. Chinese Journal of Parasitology and Parasitic Diseases, 2023, 41(3): 380-383.

图/表 3

参考文献 17

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1例蓝氏贾第鞭毛虫感染者的虫种分子鉴定及基因溯源

Molecular identification and genetic tracing of Giardia lamblia isolated from an infected case

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