山西省晋中市猪刚地弓形虫的感染情况及基因型鉴定

doi:10.12140/j.issn.1000-7423.2024.03.013

中国寄生虫学与寄生虫病杂志 ›› 2024, Vol. 42 ›› Issue (3): 367-371.doi: 10.12140/j.issn.1000-7423.2024.03.013

山西省晋中市猪刚地弓形虫的感染情况及基因型鉴定

李静¹(), 杨树峰¹, 高文伟¹, 刘卿¹, 朱兴全¹^,², 郑文斌¹^,^*()

1 山西农业大学动物医学学院，太谷 030801
2 云南农业大学动物医学院，云南省高校兽医公共卫生重点实验室，昆明 650201

收稿日期:2023-11-24 修回日期:2024-02-01 出版日期:2024-06-30 发布日期:2024-07-16
通讯作者: *郑文斌，（1991—），男，博士，副教授，从事重要动物寄生虫和人兽共患寄生虫的感染与致病机制等研究。E-mail：wenbinzheng1@126.com
作者简介:李静（2000—），女，硕士研究生，从事兽医寄生虫病学研究。E-mail：lijing1127x@163.com
基金资助:
山西农业大学动物医学学院本科生科研创新训练计划(J202111315);山西省高等学校大学生创新创业训练计划(20220147);国家重点研发计划(2021YFC2300800);国家重点研发计划(2021YFC2300802);山西农业大学高层次人才科研专项项目(2021XG001)

Infection status and genotype identification of Toxoplasma gondii in pigs in Jinzhong City, Shanxi Province

LI Jing¹(), YANG Shufeng¹, GAO Wenwei¹, LIU Qing¹, ZHU Xingquan¹^,², ZHENG Wenbin¹^,^*()

1 College of Veterinary Medicine, Shanxi Agricultural University, Taigu 030801, China
2 Key Laboratory of Veterinary Public Health of Higher Education of Yunnan Province, College of Veterinary Medicine, Yunnan Agricultural University, Kunming 650201, China

Received:2023-11-24 Revised:2024-02-01 Online:2024-06-30 Published:2024-07-16
Supported by:
Student Research and Innovation Training Program of the College of Veterinary Medicine, Shanxi Agricultural University(J202111315);College Students’ Innovative Entrepreneurial Training of Shanxi Higher Education Institution(20220147);National Key Research and Development Program of China(2021YFC2300800);National Key Research and Development Program of China(2021YFC2300802);Special Research Fund of Shanxi Agricultural University for High-level Talents(2021XG001)

摘要/Abstract

摘要：

目的了解山西省晋中市猪刚地弓形虫的感染情况和基因型现状，为猪弓形虫病的防控提供依据。方法 2021年从山西省晋中市太谷区、平遥县、昔阳县采集猪淋巴组织和肌肉组织，用动物组织提取试剂盒提取样品基因组DNA，半巢式PCR扩增弓形虫B1基因并测序。运用聚合酶链反应-限制性片段长度多态性（PCR-RFLP）方法，扩增弓形虫阳性样品和8个参考虫株的SAG1、5'-SAG2、3'-SAG2、alter. SAG2、SAG3、BTUB、GRA6、c22-8、c29-2、L358、PK1和Apico等12个位点，用限制性内切酶对扩增片段进行酶切，将弓形虫阳性样品的酶切产物与8个弓形虫国际参考虫株进行对比，通过条带大小确定基因型。结果山西省晋中市130只猪共检出37份弓形虫阳性样品，阳性率为28.5%（37/130）。其中，太谷区阳性率为32.7%（18/55），平遥县阳性率为26.0%（13/50），昔阳县阳性率为24.0%（6/25）。猪淋巴组织的弓形虫阳性率为33.0%（33/100），肌肉组织的弓形虫阳性率为13.3%（4/30）。在37份弓形虫阳性组织中，1个样品成功扩增和酶切出所有位点，鉴定为ToxoDB#9；1个样品成功扩增和酶切出8个位点，疑似ToxoDB#9、ToxoDB#20或ToxoDB#137；3个样品成功扩增和酶切出6个位点，疑似ToxoDB#9、ToxoDB#20或ToxoDB#137；1个样品成功扩增和酶切出8个位点，疑似Type Ⅰ、ToxoDB#27或ToxoDB#35；1个样品成功扩增和酶切出6个位点，疑似Type Ⅰ、ToxoDB#24、ToxoDB#27或ToxoDB#35。结论本研究对山西省晋中市猪弓形虫感染情况进行了分子检测和基因型鉴定，阳性率较高，淋巴组织阳性率高于肌肉组织，PCR-RFLP鉴定出ToxoDB#9基因型和疑似ToxoDB#9、Type Ⅰ基因型样品。

关键词: 山西省, 猪, 弓形虫, 分子检测, 基因型鉴定

Abstract:

Objective To understand the genotype characteristics of Toxoplasma gondii in Jinzhong City, Shanxi Province, and provide a basis for the prevention and control of toxoplasmosis in pigs. Methods In 2021, pig lymphatic and muscle tissues were collected from Taigu District, Pingyao County, and Xiyang County in Jinzhong City, Shanxi Province. The genomic DNA of the samples was extracted using an animal tissue extraction kit, and the B1 gene of T. gondii was amplified and sequenced using semi-nested PCR. Using the polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) method, 12 loci including SAG1, 5'-SAG2, 3'-SAG2, alter. SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico were amplified from positive samples and 8 reference strains of T. gondii. The amplified fragments were digested using restriction endonucleases, and the digested products were compared with 8 international reference strains of T. gondii. Genotypes were determined by band size. Results A total of 37 positive samples of T. gondii were detected in 130 pigs in Jinzhong City, Shanxi Province, with a positive rate of 28.5% (37/130). Among them, the positive rate in Taigu District is 32.7% (18/55), Pingyao County is 26.0% (13/50), and Xiyang County is 24.0% (6/25). The positive rate of T. gondii in pig lymphoid tissue is 33.0% (33/100), and the positive rate of T. gondii in muscle tissue is 13.3% (4/30). Out of 37 positive tissues for T. gondii, one sample was successfully amplified and all loci were cleaved, identified as ToxoDB#9; One sample successfully amplified and cleaved 8 sites, suspected to be ToxoDB#9, ToxoDB#20, or ToxoDB#137; Three samples were successfully amplified and enzyme cleaved to 6 sites, suspected to be ToxoDB#9, ToxoDB#20, or ToxoDB#137; One sample successfully amplified and cleaved 8 sites, suspected to be Type I or ToxoDB#27 or ToxoDB#35; One sample successfully amplified and cleaved 6 loci, suspected to be Type I or ToxoDB#24, ToxoDB#27, or ToxoDB#35. Conclusion This study conducted molecular detection and genotype identification of T. gondii infection in pigs in Jinzhong City, Shanxi Province. The positivity rate was high, and the positivity rate in lymphoid tissue was higher than in muscle tissue. ToxoDB#9 genotype and suspected ToxoDB#9 or Type Ⅰ genotype samples were identified by PCR-RFLP.

Key words: Shanxi Province, Pig, Toxoplasma gondii, Molecular detection, Genotyping

中图分类号:

R531.8

李静, 杨树峰, 高文伟, 刘卿, 朱兴全, 郑文斌. 山西省晋中市猪刚地弓形虫的感染情况及基因型鉴定[J]. 中国寄生虫学与寄生虫病杂志, 2024, 42(3): 367-371.

LI Jing, YANG Shufeng, GAO Wenwei, LIU Qing, ZHU Xingquan, ZHENG Wenbin. Infection status and genotype identification of Toxoplasma gondii in pigs in Jinzhong City, Shanxi Province[J]. Chinese Journal of Parasitology and Parasitic Diseases, 2024, 42(3): 367-371.

图/表 3

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分离株 Isolate	宿主 Host	分离地区 Location	SAG1	5' + 3' SAG2	Alter. SAG2	SAG3	BTUB	GRA6	c22-8	c29-2	L358	PK1	Apico	基因型 Genotype
24	猪 pig	太谷区 Taigu District	u-1	Ⅱ	Ⅱ	Ⅲ	Ⅲ	Ⅱ	Ⅱ	Ⅲ	Ⅱ	Ⅱ	Ⅰ	ToxoDB#9
1	猪 pig	太谷区 Taigu District	u-1	Ⅱ	Ⅱ	Ⅲ	N	N	N	N	Ⅱ	N	N	ToxoDB#9/ToxoDB#20/ToxoDB#137
4	猪 pig	太谷区 Taigu District	u-1	Ⅱ	Ⅱ	Ⅲ	N	N	N	N	Ⅱ	N	N	ToxoDB#9/ToxoDB#20/ToxoDB#137
10	猪 pig	太谷区 Taigu District	u-1	Ⅱ	Ⅱ	Ⅲ	Ⅲ	N	Ⅱ	N	Ⅱ	N	N	ToxoDB#9/ToxoDB#20/ToxoDB#137
11	猪 pig	太谷区 Taigu District	N	Ⅱ	N	Ⅲ	Ⅲ	Ⅱ	Ⅱ	N	N	N	N	ToxoDB#9/ToxoDB#20/ToxoDB#137
28	猪 pig	太谷区 Taigu District	N	Ⅰ	Ⅰ	Ⅰ	N	N	Ⅰ	N	Ⅰ	N	N	Type Ⅰ/ToxoDB#24/ToxoDB#27/ToxoDB#35
49	猪 pig	平遥县 Pingyao County	N	N	Ⅰ	Ⅰ	Ⅲ	N	N	N	Ⅰ	N	Ⅰ	无法确认 Indeterminacy
63	猪 pig	平遥县 Pingyao County	N	N	Ⅰ	Ⅲ	N	N	Ⅰ	Ⅰ	Ⅰ	Ⅰ	Ⅰ	无法确认 Indeterminacy
97	猪 pig	昔阳县 Xiyang County	N	Ⅰ	Ⅰ	Ⅲ	N	N	N	N	Ⅰ	Ⅰ	Ⅰ	无法确认 Indeterminacy
108	猪 pig	太谷区 Taigu District	N	Ⅱ	N	Ⅲ	N	Ⅰ	N	N	Ⅰ	N	Ⅰ	无法确认 Indeterminacy
110	猪 pig	太谷区 Taigu District	N	Ⅰ	N	Ⅲ	Ⅰ	Ⅱ	I	N	Ⅰ	N	Ⅱ	无法确认 Indeterminacy
111	猪 pig	太谷区 Taigu District	N	Ⅰ	Ⅰ	Ⅰ	Ⅰ	Ⅰ	Ⅰ	N	N	Ⅰ	N	Type Ⅰ/ToxoDB#27/ToxoDB#35
112	猪 pig	太谷区 Taigu District	N	Ⅰ	N	Ⅲ	N	N	N	Ⅱ	Ⅰ	Ⅰ	N	无法确认Indeterminacy

山西省晋中市猪刚地弓形虫的感染情况及基因型鉴定

Infection status and genotype identification of Toxoplasma gondii in pigs in Jinzhong City, Shanxi Province

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