1例蓝氏贾第鞭毛虫感染者的病原诊断与遗传进化分析

doi:10.12140/j.issn.1000-7423.2026.03.020

中国寄生虫学与寄生虫病杂志 ›› 2026, Vol. 44 ›› Issue (3): 448-453.doi: 10.12140/j.issn.1000-7423.2026.03.020

1例蓝氏贾第鞭毛虫感染者的病原诊断与遗传进化分析

余树坤¹()(), 刘浪¹, 崔紫妍¹, 谭雅心¹, 黄顺楷², 陶志阳¹^,^*()()

¹ 武汉市东西湖区疾病预防控制中心，湖北武汉 430040
² 广州市金圻睿生物科技有限责任公司，广东广州 510700

收稿日期:2025-11-21 修回日期:2026-01-20 出版日期:2026-06-30 发布日期:2026-06-12
通讯作者: *陶志阳（ORCID：0009-0007-0236-0404），男，主任医师，从事疾病预防与控制专业。E-mail：517229850@qq.com
作者简介:余树坤（ORCID：0009-0009-1657-8978），男，硕士，副主任技师，从事病原微生物检验与分子诊断工作。E-mail：411279175@qq.com
第一联系人：作者贡献
余树坤负责实验设计、撰写论文，刘浪、崔紫妍、谭雅心负责病例临床资料的收集及粪便样本检测；黄顺楷负责测序数据分析；陶志阳负责指导论文撰写。
基金资助:
武汉中青年医学骨干人才培养工程(武卫通〔2020〕55号);湖北省卫生健康委员会联合基金项目(WJ2019H406)

Pathogenic diagnosis and phylogenetic analysis of Giardia lamblia from an infected case

YU Shukun¹()(), LIU Lang¹, CUI Ziyan¹, TAN Yaxin¹, HUANG Shunkai², TAO Zhiyang¹^,^*()()

¹ Disease Prevention and Control Center of Dongxihu District, Wuhan 430040, Hubei, China
² Guangzhou Jinqi Rui Biotechnology Co., Ltd., Guangzhou 510700, Guangdong, China

Received:2025-11-21 Revised:2026-01-20 Online:2026-06-30 Published:2026-06-12
Contact: *E-mail：517229850@qq.com
Supported by:
Wuhan Middle and Young Medical Backbone Talent Training Project (〔2025〕55);Hubei Provincial Health Commission Joint Fund Project(WJ2019H406)

摘要/Abstract

摘要：

为明确1例腹泻患者感染的蓝氏贾第鞭毛虫（简称“贾第虫”）的感染来源和遗传特征，收集该患者的新鲜粪样，综合采用镜检、实时荧光定量PCR和巢式PCR方法进行病原检测，对小亚基核糖体RNA（SSU rRNA）、磷酸丙糖异构酶（tpi）、谷氨酸脱氢酶（gdh）、β-贾第素（bg）和翻译延伸因子1α（ef1α）进行宏基因组测序并分析，通过序列比对及系统进化树构建分析，开展多位点基因分型与遗传进化研究。镜检与实时荧光定量PCR均确认贾第虫感染。巢式PCR中仅gdh和ef1α基因扩增成功，宏基因组测序成功检测到SSU rRNA基因序列。多位点基因分型显示，本研究获得的分离株DXH001为集聚体B型，未发现混合感染。系统发育分析表明，DXH001分离株的gdh基因与比利时人源株EU847734的序列一致性达100%，属BⅣ亚型；ef1α基因与澳大利亚人源分离株HQ179598的序列一致性达98.43%，SSU rRNA基因与美国人源株U09491的序列一致性达90.45%，均聚类于B型分支。SSU rRNA基因与人源参考序列U09491相比存在35个碱基替换和5个缺失，其序列差异可能主要与样本中目标序列丰度较低，导致测序数据量不足有关；gdh与ef1α基因的变异较为保守，未引起重要功能改变。本研究通过多位点基因分析鉴定DXH001株为蓝氏贾第鞭毛虫集聚体BⅣ亚型，提示该亚型存在跨地域传播风险。宏基因组测序在低丰度靶标检测中展现出较高灵敏度，可作为常规分子分型的有效补充。

关键词: 蓝氏贾第鞭毛虫, 腹泻感染, 分子病原学, 系统发育树, 变异位点

Abstract:

To clarify the infection source and genetic characteristics of Giardia lamblia isolated from a diarrheal patient, fresh fecal sample of the patient was collected. Microscopy, real-time fluorescent quantitative PCR and nested PCR were comprehensively applied for pathogen detection. Metagenomic sequencing and analysis were performed targeting the small subunit ribosomal RNA (SSU rRNA), triosephosphate isomerase (tpi), glutamate dehydrogenase (gdh), β-giardin (bg) and translation elongation factor 1α (ef1α). Multilocus genotyping and genetic evolution analysis were carried out via sequence alignment and phylogenetic tree construction. Both microscopy and real-time fluorescent quantitative PCR confirmed Giardia infection. Only the gdh and ef1α genes were successfully amplified by nested PCR, and the SSU rRNA gene sequence was successfully detected by metagenomic sequencing. Multilocus genotyping showed that the isolate DXH001 obtained in this study belonged to assemblage B, and no mixed infection was detected. Phylogenetic analysis indicated that the gdh gene of isolate DXH001 shared 100% sequence identity with the human-derived strain EU847734 from Belgium, belonging to subtype BⅣ; the ef1α gene shared 98.43% sequence identity with the human-derived isolate HQ179598 from Australia, and the SSU rRNA gene shared 90.45% sequence identity with the human-derived strain U09491 from the United States, both clustered into the assemblage B branch. Compared with the human-derived reference sequence U09491, the SSU rRNA gene had 35 base substitutions and 5 deletions, and the sequence difference was mainly related to the insufficient sequencing data volume caused by the low abundance of the target sequence in the sample; the variations of gdh and ef1α genes were relatively conservative, which did not cause significant functional changes. In this study, the isolate DXH001 was identified as G. lamblia assemblage BⅣ via multilocus genetic analysis, suggesting the trans-regional transmission risk of this subtype. Metagenomic sequencing shows high sensitivity in low-abundance target detection, which can serve as an effective supplement for conventional molecular genotyping.

Key words: Giardia lamblia, Diarrheal infection, Molecular aetiology, Phylogenetic tree, Variable loci

中图分类号:

R531.73

余树坤, 刘浪, 崔紫妍, 谭雅心, 黄顺楷, 陶志阳. 1例蓝氏贾第鞭毛虫感染者的病原诊断与遗传进化分析[J]. 中国寄生虫学与寄生虫病杂志, 2026, 44(3): 448-453.

YU Shukun, LIU Lang, CUI Ziyan, TAN Yaxin, HUANG Shunkai, TAO Zhiyang. Pathogenic diagnosis and phylogenetic analysis of Giardia lamblia from an infected case[J]. Chinese Journal of Parasitology and Parasitic Diseases, 2026, 44(3): 448-453.

图/表 5

表1

图1

图2

图3

图4

参考文献 16

[1]	王丹, 贺志权, 刘颖, 等. 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 D, He ZQ, Liu Y, et al. Molecular identification and genetic tracing of Giardia lamblia isolated from an infected case[J]. Chin J Parasitol Parasit Dis, 2023, 41(3): 380-383. (in Chinese)
[2]	Fink MY, Shapiro D, Singer SM. Giardia Lamblia: laboratory maintenance, lifecycle induction, and infection of murine models[J]. Curr Protoc Microbiol, 2020, 57(1): e102.
[3]	Conners EE, Miller AD, Balachandran N, et al. Giardiasis outbreaks - United States, 2012-2017[J]. MMWR Morb Mortal Wkly Rep, 2021, 70(9): 304-307.
[4]	Zhang XP, Dan JM, Wang LQ, et al. High genetic diversity of Giardia duodenalis assemblage E in Chinese dairy cattle[J]. Infect Genet Evol, 2021, 92: 104912.
[5]	Dougherty M, Bartelt LA. Giardia and growth impairment in children in high-prevalence settings: consequence or co-incidence?[J]. Curr Opin Infect Dis, 2022, 35(5): 417-423.
[6]	Traub RJ, Monis PT, Robertson I, et al. Epidemiological and molecular evidence supports the zoonotic transmission of Giardia among humans and dogs living in the same community[J]. Parasitology, 2004, 128(Pt 3): 253-262.
[7]	Read CM, Monis PT, Andrew Thompson RC. Discrimination of all genotypes of Giardia duodenalis at the glutamate dehydrogenase locus using PCR-RFLP[J]. Infect Genet Evol, 2004, 4(2): 125-130.
[8]	Hopkins RM, Meloni BP, Groth DM, et al. Ribosomal RNA sequencing reveals differences between the genotypes of Giardia isolates recovered from humans and dogs living in the same locality[J]. J Parasitol, 1997, 83(1): 44-51.
[9]	Sulaiman IM, Fayer R, Bern C, et al. Triosephosphate isomerase gene characterization and potential zoonotic transmission of Giardia duodenalis[J]. Emerg Infect Dis, 2003, 9(11): 1444-1452.
[10]	Cacciò SM, de Giacomo M, Pozio E. Sequence analysis of the beta-giardin gene and development of a polymerase chain reaction-restriction fragment length polymorphism assay to genotype Giardia duodenalis cysts from human faecal samples[J]. Int J Parasitol, 2002, 32(8): 1023-1030.
[11]	Fletcher SM, Stark D, Harkness J, et al. Enteric protozoa in the developed world: a public health perspective[J]. Clin Microbiol Rev, 2012, 25(3): 420-449.
[12]	蔡伟龙, 李娜, 冯耀宇, 等. 牛十二指肠贾第虫的分子流行病学研究进展[J]. 畜牧兽医学报, 2021, 52(2): 300-310.
	Cai WL, Li N, Feng YY, et al. Advances in molecular epidemiology of Giardia duodenalis from cattle in China[J]. Acta Vet Zootechnica Sin, 2021, 52(2): 300-310. (in Chinese)
[13]	王斌. 上海地区动物园野生动物贾第虫感染调查及其分子特征研究[D]. 遵义: 遵义医学院, 2015.
	Wang B. Investigation of Giardia infection in wild animals in zoos in Shanghai and study on their molecular characteristics[D]. Zunyi: Zunyi Medcial College, 2015. (in Chinese)
[14]	Ryan U, Zahedi A. Molecular epidemiology of giardiasis from a veterinary perspective[J]. Adv Parasitol, 2019, 106: 209-254.
[15]	Hashemi-Hafshejani S, Meamar AR, Moradi M, et al. Multilocus sequence typing of Giardia duodenalis genotypes circulating in humans in a major metropolitan area[J]. Front Med, 2022, 9: 976956.
[16]	温少芳, 卢思奇. 蓝氏贾第鞭毛虫遗传多样性研究进展[J]. 国际医学寄生虫病杂志, 2006, 33(1): 44-48.
	Wen SF, Lu SQ. Advance in genetic diversity of Giardia lamblia[J]. Int J Med Parasit Dis, 2006, 33(1): 44-48. (in Chinese)

基因名称	引物序列（5'→3'）	片段大小/bp
SSU rRNA	F1：CATCCGGTCGATCCTGCC	292
	R1：AGTCGAACCCTGATTCTCCGCCCAGG	292
	F2：GACGCTCTCCCCAAGGAC	174
	R2：CTGCGTCACGCTGCTCG	174
gdh	F1：TTCCGTRTYCAGTACAACTC	829
	R1：ACCTCGTTCTGRGTGGCGCA3	829
	F2：ATGACYGAGCTYCAGAGGCACGT	432
	R2：GTGGCGCARGGCATGATGCA	432
tpi	F1：AAATIATGCCTGCTCGTCG	605
	R1：CAAACCTTITCCGCAAACC	605
	F2：CCCTTCATCGGIGGTAACTT	530
	R2：GTGGCCACCACICCCGTGCC	530
bg	F1：AAGCCCGACGACCTCACCCGCAGTGC	753
	R1：GAGGCCGCCCTGGATCTTCGAGACGAC	753
	F2：CATAACGACGCCATCGCGGCTCTCAGGAA	384
	R2：GAGGCCGCCCTGGATCTTCGAGACGAC	384
ef1α	F：GCCGAGGAGTTCGACTACATC	191
ef1α	R：GACGCCSGAGATCTTGTAGAC	191

1例蓝氏贾第鞭毛虫感染者的病原诊断与遗传进化分析

Pathogenic diagnosis and phylogenetic analysis of Giardia lamblia from an infected case

RichHTML

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

图/表 5

参考文献 16

相关文章 15

编辑推荐

Metrics

本文评价

[1]	乌云其木格, 道嘉昊, 蔡然, 付松青, 阮星宇, 单雨玮, 付雅婷, 胡薇. 微小隐孢子虫、蓝氏贾第鞭毛虫和牛艾美球虫胶体金免疫层析快速检测方法的建立[J]. 中国寄生虫学与寄生虫病杂志, 2026, 44(3): 429-436.
[2]	彭晓雪, 王旭, 简金花, 沈玉娟. 我国人群蓝氏贾第鞭毛虫感染及食源性污染现状[J]. 中国寄生虫学与寄生虫病杂志, 2025, 43(2): 274-281.
[3]	闵向阳, 冯萌, 赵旭鸿, 翁文浩, 李汉华. 1例蓝氏贾第鞭毛虫感染的诊断[J]. 中国寄生虫学与寄生虫病杂志, 2024, 42(2): 272-274.
[4]	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.
[5]	朱名超, 朱娅. HIV合并人芽囊原虫和蓝氏贾第鞭毛虫感染1例[J]. 中国寄生虫学与寄生虫病杂志, 2022, 40(3): 315-318.
[6]	李淑凝, 李汶霖, 沈海娥, 王洋, 田喜凤. 蓝氏贾第鞭毛虫滋养体体外诱导中性粒细胞胞外诱捕网的形成[J]. 中国寄生虫学与寄生虫病杂志, 2021, 39(4): 455-460.
[7]	黎敏, 骆清清, 房辉, 曹晓琴. 花椒乙醇提取物体外抗蓝氏贾第鞭毛虫的作用[J]. 中国寄生虫学与寄生虫病杂志, 2021, 39(3): 359-364.
[8]	于铭川, 杨忠委, 王华然, 师丹阳, 周树青, 尹静, 孙树民. 基于LAMP微流控芯片检测蓝氏贾第鞭毛虫方法的建立[J]. 中国寄生虫学与寄生虫病杂志, 2021, 39(3): 402-405.
[9]	袁义雪, 郑国侠, 王丹丹, 专行, 杜萌, 王云华. 蓝氏贾第鞭毛虫核苷二磷酸激酶基因的克隆与表达[J]. 中国寄生虫学与寄生虫病杂志, 2019, 37(2): 190-193.
[10]	曹胜魁, 姜岩岩, 袁忠英, 徐梦, 曹建平, 杨益超, 沈玉娟. 广西宾阳县和灵山县水源隐孢子虫和蓝氏贾第鞭毛虫污染调查[J]. 中国寄生虫学与寄生虫病杂志, 2018, 36(6): 597-601.
[11]	徐宁, 尹建海, 沈玉娟, 刘华, 曹建平. 隐孢子虫和蓝氏贾第鞭毛虫分子流行病学研究进展[J]. 中国寄生虫学与寄生虫病杂志, 2018, 36(6): 661-666.
[12]	袁义雪1，郑国侠2，王丹丹1，专行1，杜萌1，王云华1*. 蓝氏贾第鞭毛虫核苷二磷酸激酶基因的克隆与表达[J]. 中国寄生虫学与寄生虫病杂志, 2018, 36(2): 190-192.
[13]	章乐生, 孙磊, 胡媛, 巩文词, 王燕娟, 沈玉娟, 徐馀信, 汪天平, 曹建平. 蓝氏贾第鞭毛虫单克隆抗体的制备及鉴定[J]. 中国寄生虫学与寄生虫病杂志, 2017, 35(2): 160-163.
[14]	俞英昉，吴秀萍，储言红，陈家旭，田利光*. 安徽农村地区HIV感染者和幼儿园儿童蓝氏贾第鞭毛虫感染情况及其基因型[J]. 中国寄生虫学与寄生虫病杂志, 2016, 34(6): 11-537-541.
[15]	魏超君1，贾彦娟1，徐辉1，吴玲1，卢思奇2，李永红1*. 蓝氏贾第鞭毛虫α-8贾第素特异性优势抗原表位肽的抗原性分析[J]. 中国寄生虫学与寄生虫病杂志, 2016, 34(5): 1-389-393.