Expert consensus on species identification of Trichinella spp. by nucleic acid method (2025)

doi:10.12140/j.issn.1000-7423.2025.05.002

Abstract

Abstract:

The larvae of Trichinella spp. nematodes parasitize in humans and various vertebrates, which seriously endangers people’s health and animal meat food safety. There are numerous species within the Trichinella genus; and the infectivity and pathogenicity of Trichinella spp. larvae are obviously different. In recent years, new Trichinella species have been discovered consecutively. The available pathogenic examination methods(direct microscopy of biopsy muscles and artificial digestion) could not distinguish the Trichinella species. To provide reference for Trichinella taxonomic researches and the accurate diagnosis of trichinellosis, this consensus elaborate on the nucleic acid methods for the identification of Trichinella spp. based on the latest domestic and international research progress.

Key words: Trichinella, Trichinellosis, Species identification, Multiplex PCR, Restriction fragment length polymorphism

CLC Number:

R382.312

ZHANG Xi, CUI Jing, LIU Ruodan, LONG Shaorong, CHEN Muxin, CHEN Jiaxu, FU Baoquan, LIU Xiaolei, LIU Mingyuan, ZHANG Yalan, ZHANG Hongwei, WANG Zhongquan. Expert consensus on species identification of Trichinella spp. by nucleic acid method (2025)[J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2025, 43(5): 607-613.

Figures/Tables 6

References 25

[1]	中华人民共和国卫生部. 旋毛虫病的诊断: WS/T 369—2012[S]. 北京: 中国标准出版社, 2012: 1-16.
	Ministry of Health of the People’s Republic of China. Diagnosis of trichinellosis: WS/T 369-2012[S]. Beijing: Standards Press of China, 2012: 1-16. (in Chinese)
[2]	Crisóstomo-Jorquera V, Landaeta-Aqueveque C. The genus Trichinella and its presence in wildlife worldwide: A review[J]. Transbound Emerg Dis, 2022, 69(5): e1269-e1279.
[3]	Miguez S, Moreno María A, Fariña FA, et al. Understanding the global connection: Investigating the association between pork meat exports and trichinellosis[J]. Vet Parasitol, 2025, 338: 110509.
[4]	Zhang XZ, Wang ZQ, Cui J. Epidemiology of trichinellosis in the People’s Republic of China during 2009-2020[J]. Acta Trop, 2022, 229: 106388.
[5]	王中全, 崔晶. 旋毛虫属分类的研究进展[J]. 中国寄生虫学与寄生虫病杂志, 2002, 20(5): 310-314.
	Wang ZQ, Cui J. Recent progress in the taxonomy of the genus Trichinella[J]. Chin J Parasitol Parasit Dis, 2002, 20(5): 310-314. (in Chinese)
[6]	Zarlenga D, Thompson P, Pozio E. Trichinella species and genotypes[J]. Res Vet Sci, 2020, 133: 289-296.
[7]	国家市场监督管理总局, 国家标准化管理委员会. 旋毛虫诊断技术: GB/T 18642—2021[S]. 北京: 中国标准出版社, 2021: 1-28.
	State Administration for Market Regulation, National Standardization Administration. Diagnostic techniques for Trichinella spp.: GB/T 18642-2021[S]. Beijing: Standards Press of China, 2021: 1-28. (in Chinese)
[8]	Pozio E, Zarlenga D. International Commission on Trichinellosis: Recommendations for genotyping Trichinella muscle stage larvae[J]. Food Waterborne Parasitol, 2019, 15: e00033.
[9]	崔晶, 王中全. 旋毛虫与旋毛虫病的若干术语释义[J]. 中国寄生虫学与寄生虫病杂志, 2024, 42(4): 427-432.
	Cui J, Wang ZQ. Interpretations of some terminology of Trichinella and trichinellosis[J]. Chin J Parasitol Parasit Dis, 2024, 42(4): 427-432. (in Chinese)
[10]	Pozio E. Scientific achievements of the last 60 years: From a single to a multispecies concept of the genus Trichinella[J]. Vet Parasitol, 2021, 297: 109042.
[11]	Barlow A, Roy K, Hawkins K, et al. A review of testing and assurance methods for Trichinella surveillance programs[J]. Food Waterborne Parasitol, 2021, 24: e00129.
[12]	Bilska-Zając E, Thompson P, Rosenthal B, et al. Infection, genetics, and evolution of Trichinella: Historical insights and applications to molecular epidemiology[J]. Infect Genet Evol, 2021, 95: 105080.
[13]	Malone CJ, Oksanen A, Mukaratirwa S, et al. From wildlife to humans: The global distribution of Trichinella species and genotypes in wildlife and wildlife-associated human trichinellosis[J]. Int J Parasitol Parasites Wildl, 2024, 24: 100934.
[14]	Pozio E, Owen IL, Rosa GL, et al. Trichinella papuae n. sp. (Nematoda), a new non-encapsulated species from domestic and sylvatic swine of Papua New Guinea[J]. Int J Parasitol, 1999, 29(11): 1825-1839.
[15]	Pozio E, Foggin CM, Marucci G, et al. Trichinella zimbabwensis n.sp. (Nematoda), a new non-encapsulated species from crocodiles (Crocodylus niloticus) in Zimbabwe also infecting mammals[J]. Int J Parasitol, 2002, 32(14): 1787-1799.
[16]	Krivokapich SJ, Pozio E, Gatti GM, et al. Trichinella patagoniensis n. sp. (Nematoda), a new encapsulated species infecting carnivorous mammals in South America[J]. Int J Parasitol, 2012, 42(10): 903-910.
[17]	Sharma R, Thompson PC, Hoberg EP, et al. Hiding in plain sight: Discovery and phylogeography of a cryptic species of Trichinella (Nematoda: Trichinellidae) in wolverine (Gulo gulo)[J]. Int J Parasitol, 2020, 50(4): 277-287.
[18]	Zarlenga DS, Chute MB, Martin A, et al. A multiplex PCR for unequivocal differentiation of all encapsulated and non-encapsulated genotypes of Trichinella[J]. Int J Parasitol, 1999, 29(11): 1859-1867.
[19]	崔晶, 赵桂花, 王中全, 等. 应用多重PCR技术鉴定我国6个旋毛虫地理株的研究[J]. 中国寄生虫学与寄生虫病杂志, 2008, 26(2): 86-89.
	Cui J, Zhao GH, Wang ZQ, et al. Identification of six Trichinella isolates from China by multiplex PCR[J]. Chin J Parasitol Parasit Dis, 2008, 26(2): 86-89. (in Chinese)
[20]	Jiang P, Zhang X, Wang LA, et al. Survey of Trichinella infection from domestic pigs in the historical endemic areas of Henan Province, central China[J]. Parasitol Res, 2016, 115(12): 4707-4709.
[21]	Fu BQ, Liu MY, Yao CY, et al. Species identification of Trichinella isolates from China[J]. Vet Parasitol, 2009, 159(3/4): 214-217.
[22]	Zhang X, Han LL, Hong X, et al. Genotyping and phylogenetic position of Trichinella spiralis isolates from different geographical locations in China[J]. Front Genet, 2019, 10: 1093.
[23]	Pozio E, Pagani P, Marucci G, et al. Trichinella britovi etiological agent of sylvatic trichinellosis in the Republic of Guinea (West Africa) and a re-evaluation of geographical distribution for encapsulated species in Africa[J]. Int J Parasitol, 2005, 35(9): 955-960.
[24]	Nagano I, Wu Z, Matsuo A, et al. Identification of Trichinella isolates by polymerase chain reaction: Restriction fragment length polymorphism of the mitochondrial cytochrome c-oxidase subunit I gene[J]. Int J Parasitol, 1999, 29(7): 1113-1120.
[25]	Lobanov VA, Konecsni KA, Brad Scandrett W, et al. Identification of Trichinella taxa by ITS-1 amplicon next-generation sequencing with an improved resolution for detecting underrepresented genotypes in mixed natural infections[J]. Parasit Vectors, 2023, 16(1): 466.

旋毛虫种/基因型	ISS编码	宿主来源	国家	分离年份
旋毛虫（T. spiralis，T1）	ISS3	野猪（Sus scrofa）	波兰	1960
乡土旋毛虫（T. nativa，T2）	ISS10	北极熊（Ursus maritimus）	挪威	1984
布氏旋毛虫（T. britovi，T3）	ISS120	赤狐（Vulpes vulpes）	意大利	1988
伪旋毛虫（T. pseudospiralis，T4）	ISS13	浣熊（Procyon lotor）	俄罗斯	1972
穆氏旋毛虫（T. murrelli，T5）	ISS417	草原狼（Canis latrans）	美国	1989
基因型T6（Trichinella，T6）	ISS34	灰熊（Ursus arctos horribilis）	美国	1983
纳氏旋毛虫（T. nelsoni，T7）	ISS37	疣猪（Phacochoerus aethiopicus）	坦桑尼亚	1982
基因型T8（Trichinella，T8）	ISS272	非洲狮（Panthera leo）	纳米比亚	1992
基因型T9（Trichinella，T9）	ISS409	貉（Nyctereutes procyonoides）	日本	1984
巴布亚旋毛虫（T. papuae，T10）	ISS572	野猪（Sus scrofa）	巴布亚新几内亚	1997
津巴布韦旋毛虫（T. zimbabwensis，T11）	ISS1029	尼罗鳄（Crocodylus niloticus）	津巴布韦	1995
巴塔哥尼亚旋毛虫（T. patagoniensis，T12）	ISS2496	美洲狮（Puma concolor）	阿根廷	2008
羌查旋毛虫（T. chanchalensis，T13）		狼獾（Gulo gulo）	加拿大	2020

目的基因	引物编号	引物序列（5'→3'）
ESⅤ	cp-Ⅰ.F	GTTCCATGTGAACAGCAGT
ESⅤ	cp-Ⅰ.R	CGAAAACATACGACAACTGC
ITS1	cp-Ⅱ.F	GCTACATCCTTTTGATCTGTT
	cp-Ⅱ.R	AGACACAATATCAACCACAGTACA
	cp-Ⅲ.F	GCGGAAGGATCATTATCGTGTA
	cp-Ⅲ.R	TGGATTACAAAGAAAACCATCACT
ITS2	cp-Ⅳ.F	GTGAGCGTAATAAAGGTGCAG
	cp-Ⅳ.R	TTCATCACACATCTTCCACTA
	cp-Ⅴ.F	CAATTGAAAACCGCTTAGCGTGTTT
	cp-Ⅴ.R	TGATCTGAGGTCGACATTTCC

旋毛虫虫种/基因型	目的基因（条带大小/bp）
旋毛虫	ESⅤ（173）
乡土旋毛虫/羌查旋毛虫	ESⅤ（129）
布氏旋毛虫/T8/T9	ESⅤ（129）、ITS1（253）
伪旋毛虫	ESⅤ（292~360）
穆氏旋毛虫	ESⅤ（129）、ITS2（316）
T6	ESⅤ（129）、ITS1（210）
纳氏旋毛虫	ESⅤ（155）、ITS2（404）
巴布亚旋毛虫	ESⅤ（240）
巴塔哥尼亚旋毛虫	ESⅤ（264）
羌查旋毛虫	ESⅤ（127）

目的基因	引物序列（5'→3'）	区分虫种
cytb^[17]	F：ATACGAGAAGTAAAATTTGG	乡土旋毛虫/羌查旋毛虫
cytb^[17]	R：TCATTGAATAGAGTGTGGTT	乡土旋毛虫/羌查旋毛虫
ITS2^[23]	F：CCGGTGAGCGTAATAAAG	布氏旋毛虫/T8基因型
ITS2^[23]	R：TACACACAACGCAACGAT	布氏旋毛虫/T8基因型
cox1^[24]	F：TTYTGGTTYTTCGGKCACCC	布氏旋毛虫/T9基因型
cox1^[24]	R：ACGACGTAGTAGGTRTCRTG	布氏旋毛虫/T9基因型