Hostname: page-component-78c5997874-94fs2 Total loading time: 0 Render date: 2024-11-17T15:14:11.022Z Has data issue: false hasContentIssue false

Use of mitochondrial RNA genes for the differentiation of four Trichinella species by multiplex PCR amplification

Published online by Cambridge University Press:  01 June 2009

R. Blaga*
Affiliation:
University of Agricultural Sciences and Veterinary Medicine, 400372Cluj-Napoca, Romania
BaoQuan Fu
Affiliation:
Key Laboratory of Veterinary Parasitology of Gansu Province, State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, 730046Lanzhou, P.R. China
D. Le Rhun
Affiliation:
JRU BIPAR INRA AFSSA ENVA UPVM, 94703Maisons-Alfort cedex, France
E. Le Naour
Affiliation:
JRU BIPAR INRA AFSSA ENVA UPVM, 94703Maisons-Alfort cedex, France
A. Heckman
Affiliation:
JRU BIPAR INRA AFSSA ENVA UPVM, 94703Maisons-Alfort cedex, France
A. Zocevic
Affiliation:
JRU BIPAR INRA AFSSA ENVA UPVM, 94703Maisons-Alfort cedex, France
MingYuan Liu
Affiliation:
Key Laboratory of Zoonoses, Ministry of Education, Institute of Zoonoses, Jilin University, 5333 Xian Road, 130062Changchun, P.R. China
P. Boireau
Affiliation:
JRU BIPAR INRA AFSSA ENVA UPVM, 94703Maisons-Alfort cedex, France
*

Abstract

Until now, four species of the Trichinella genus have been identified in Europe: Trichinella spiralis, T. nativa, T. britovi and T. pseudospiralis. The aim of this work was to establish a sound polymerase chain reaction (PCR)-based method to differentiate these four species using mitochondrial rDNA as a reliable genetic marker and to evaluate the sensitivity of this method. Full-length DNA sequences coding for the small and large mitochondrial rRNA (mt-rrnS and mt-rrnL) of the four species are described. A multiplex PCR was designed and successfully tested on 24 European isolates. As few as one larva, or 100 pg of genomic DNA was detected, providing equivalent sensitivity to previously described PCR methods. The PCR-based method of mitochondrial rDNA amplification was thereby established as a sensitive and reproductive diagnostic method for the four European Trichinella species.

Type
Research Papers
Copyright
Copyright © Cambridge University Press 2009

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Anderson, T.J., Komuniecki, R., Komuniecki, P.R. & Jaenike, J. (1995) Are mitochondria inherited paternally in Ascaris? International Journal for Parasitology 25, 10011004.CrossRefGoogle ScholarPubMed
Anderson, T.J., Blouin, M.S. & Beech, R.N. (1998) Population biology of parasitic nematodes: applications of genetic markers. Advances in Parasitology 41, 219283.CrossRefGoogle ScholarPubMed
Appleyard, G.D., Zarlenga, D.S., Pozio, E. & Gajadhar, A.A. (1999) Differentiation of Trichinella genotypes by PCR using sequence specific primers. Journal of Parasitology 85, 556559.CrossRefGoogle ScholarPubMed
Arriaga, C., Yepes-Mulia, L., Viveros, N., Zarlenga, D.S., Lichtenfels, J.R., Benitez, E. & Ortega-Pierres, M.G. (1995) Detection of Trichinella spiralis muscle larvae in naturally-infected horses. Journal of Parasitology 81, 781783.CrossRefGoogle ScholarPubMed
Bandi, C., La Rosa, G., Bardin, M.G., Damiani, G., de Carneri, I. & Pozio, E. (1993) Arbitrarily primed polymerase chain reaction of individual Trichinella specimens. Journal of Parasitology 79, 437440.CrossRefGoogle ScholarPubMed
Beck, R., Beck, A., Lučinger, S., Florijančić, T., Bošković, I. & Marinculić, A. (2008) Trichinella pseudospiralis in pigs from Croatia. Veterinary Parasitology 159, 304307.CrossRefGoogle ScholarPubMed
Blouin, M.S. (2002) Molecular prospecting for cryptic species of nematodes: mitochondrial DNA versus internal transcribed spacer. International Journal for Parasitology 32, 527531.CrossRefGoogle ScholarPubMed
Borsuk, P., Moskwa, B., Pastusiak, K. & Cabaj, W. (2003) Molecular identification of Trichinella spiralis and Trichinella britovi by diagnostic multiprimer large mitochondrial rRNA amplification. Parasitology Research 91, 374377.CrossRefGoogle ScholarPubMed
Britov, V.A. & Boev, S.N. (1972) Taxonomic rank of various strains of Trichinella and their circulation in nature. Vestink Akademii Nauk SSSR 28, 2732.Google Scholar
Brown, W.M. (1985) The mitochondrial genome of animals. pp. 95130in MacIntyre, R.J. (Ed.) Molecular evolutionary genetics. New-York, Plenum Press.CrossRefGoogle Scholar
Community Reference Laboratory for Parasites (2007) Guidelines for the identification of Trichinella muscle stage larvae at the species level by multiplex PCR. Available athttp://www.iss.it/binary/crlp/cont/PCR%20method%20WEB%20SITE.1177083731.pdf (accessed at 1st of December 2008).Google Scholar
Corpet, F. (1988) Multiple sequence alignment with hierarchical clustering. Nucleic Acids Research 16, 1088110890.CrossRefGoogle ScholarPubMed
De Bruyne, A., Yera, H., Le Guerhier, F., Boireau, P. & Dupouy-Camet, J. (2005) Simple species identification of Trichinella isolates by amplification and sequencing of the 5S ribosomal DNA intergenic spacer region. Veterinary Parasitology 132, 5761.CrossRefGoogle ScholarPubMed
Dick, T.A. & Pozio, E. (2001) Trichinella spp. and trichinellosis. pp. 380396in Samuel, W.M., Pybus, M.J. & Kocan, A.A. (Eds) Parasitic diseases of wild mammals. Ames, Iowa, Iowa State University Press.CrossRefGoogle Scholar
Dupouy-Camet, J., Robert, F., Guillou, J.P., Vallet, C., Perret, C. & Soulé, C. (1994) Identification of Trichinella isolates with random amplified polymorphic DNA markers. Parasitology Research 80, 358360.CrossRefGoogle ScholarPubMed
Gamble, H.R., Bessonov, A.S., Cuperlovic, K., Gajadhar, A.A., van Knapen, F., Noeckler, K., Schenone, H. & Zhu, X. (2000) International Commission on Trichinellosis: recommendations on methods for the control of Trichinella in domestic and wild animals intended for human consumption. Veterinary Parasitology 93, 393408.CrossRefGoogle ScholarPubMed
Garkavi, B.L. (1972) The species of Trichinella isolated from wild carnivores. Veterinariya 10, 9091.Google ScholarPubMed
Gasser, R.B., Zhu, X., Monti, J.R., Dou, L., Cai, X. & Pozio, E. (1998) PCR-SSCP of rDNA for the identification of Trichinella isolates from mainland China. Molecular and Cellular Probes 12, 2734.CrossRefGoogle ScholarPubMed
Gasser, R.B., Hu, M., El-Osta, Y.A., Zarlenga, D.S. & Pozio, E. (2005) Genetic analysis of Trichinella populations by ‘cold’ single-strand conformation polymorphism analysis. Veterinary Parasitology 132, 2326.CrossRefGoogle ScholarPubMed
Hu, M. & Gasser, R.B. (2006) Mitochondrial genomes of parasitic nematodes—progress and perspectives. Trends in Parasitology 22, 7884.CrossRefGoogle Scholar
Hu, M., Chilton, L.B. & Gasser, R.B. (2004) The mitochondrial genomics of parasitic nematodes of socio-economic importance: recent progress, and implications for population genetics and systematics. Advances in Parasitology 56, 133212.CrossRefGoogle ScholarPubMed
Kapel, C.M., Oivanen, L., La Rosa, G., Mikkonen, T. & Pozio, E. (2001) Evaluation of two PCR-based techniques for molecular epidemiology in Finland, a high-endemic area with four sympatric Trichinella species. Parasite 8, 3943.CrossRefGoogle ScholarPubMed
Krivokapich, S.J., Prous, C.L., Gatti, G.M., Confalonieri, V., Molina, V., Matarasso, H. & Guarnera, E. (2008) Molecular evidence for a novel encapsulated genotype of Trichinella from Patagonia, Argentina. Veterinary Parasitology 156, 234240.CrossRefGoogle ScholarPubMed
La Rosa, G., Marucci, G., Zarlenga, D.S. & Pozio, E. (2001) Trichinella pseudospiralis populations of the Palearctic region and their relationship with populations of the Nearctic and Australian regions. International Journal for Parasitology 31, 297305.CrossRefGoogle ScholarPubMed
Lavrov, D.V. & Brown, W.M. (2001) Trichinella spiralis mtDNA: a nematode mitochondrial genome that encodes a putative ATP8 and normally structured tRNAs and has a gene arrangement relatable to those of coelomate metazoans. Genetics 157, 621637.CrossRefGoogle Scholar
Murrell, K.D., Lichtenfels, R.J., Zarlenga, D.S. & Pozio, E. (2000) The systematics of Trichinella with a key to species. Veterinary Parasitology 93, 293307.CrossRefGoogle ScholarPubMed
Nagano, I., Wu, Z., Matsuo, I., Pozio, E. & Takahashi, Y. (1999) Identification of Trichinella genotypes by polymerase chain reaction–restriction fragment length polymorphism of mitochondrial cytochrome c oxidase subunit I gene. International Journal for Parasitology 29, 11131120.CrossRefGoogle ScholarPubMed
Nockler, K., Reckinger, S. & Pozio, E. (2006) Trichinella spiralis and Trichinella pseudospiralis mixed infection in a wild boar (Sus scrofa) of Germany. Veterinary Parasitology 137, 364368.CrossRefGoogle Scholar
Pozio, E. (2001) New patterns of Trichinella infections. Veterinary Parasitology 98, 133148.CrossRefGoogle Scholar
Pozio, E. & La Rosa, G. (2003) PCR-derived methods for the identification of Trichinella parasites from animal and human samples. Methods in Molecular Biology 216, 299309.Google ScholarPubMed
Pozio, E. & Zarlenga, D.S. (2005) Recent advances on the taxonomy, systematics and epidemiology of Trichinella. International Journal for Parasitology 35, 11911204.CrossRefGoogle ScholarPubMed
Pozio, E., Serrano, F.J., La Rosa, G., Reina, D., Perez-Martin, E. & Navarrete, I. (1997) Evidence of potential gene flow in Trichinella spiralis and in Trichinella britovi in nature. Journal of Parasitology 83, 163166.CrossRefGoogle ScholarPubMed
Rombout, Y.B., Bosch, S. & Van Der Giessen, J.W. (2001) Detection and identification of eight Trichinella genotypes by reverse line blot hybridization. Journal of Clinical Microbiology 39, 642646.CrossRefGoogle ScholarPubMed
Rosenthal, B.M., La Rosa, G., Zarlenga, D., Dunams, D., Chunyu, Y., Mingyuan, L. & Pozio, E. (2008) Human dispersal of Trichinella spiralis in domesticated pigs. Infection, Genetics, Evolution 8, 799805.CrossRefGoogle ScholarPubMed
Soulé, C., Guillou, J.P., Dupouy-Camet, J., Vallet, C. & Pozio, E. (1993) Differentiation of Trichinella isolates by polymerase chain reaction. Parasitology Research 79, 461465.CrossRefGoogle ScholarPubMed
Széll, Z., Marucci, G., Bajmóczy, E., Cséplo, A., Pozio, E. & Sréter, T. (2008) Spatial distribution of Trichinella britovi, T. pseudospiralis and T. spiralis in red foxes (Vulpes vulpes) in Hungary. Veterinary Parasitology 156, 210215.CrossRefGoogle Scholar
Tighe, P.J., Goyal, P.K., Wilson, Z.A., Wakelin, D. & Pritchard, D.I. (1994) Analysis of genetic variation in isolates of Trichinella using random amplified polymorphic DNA. Molecular and Biochemical Parasitology 63, 175178.CrossRefGoogle ScholarPubMed
van der Giessen, J.W., Fonville, M., Briels, I. & Pozio, E. (2005) Phylogenetic analysis of encapsulated and non-encapsulated Trichinella species by studying the 5S rDNA tandemly repeated intergenic region. Veterinary Parasitology 132, 5155.CrossRefGoogle ScholarPubMed
Wu, Z., Nagano, I. & Takahashi, Y. (1998) The detection of Trichinella with polymerase chain reaction (PCR) primers constructed using sequences of random amplified polymorphic DNA (RAPD) or sequences of complementary DNA encoding excretory–secretory (ES) glycoproteins. Parasitology 117, 173183.CrossRefGoogle ScholarPubMed
Wu, Z., Nagano, I., Pozio, E. & Takahashi, Y. (1999) Polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP) for the identification of Trichinella isolates. Parasitology 118, 211218.CrossRefGoogle ScholarPubMed
Zarlenga, D.S. & La Rosa, D.G. (2000) Molecular and biochemical methods for parasite differentiation within the genus Trichinella. Veterinary Parasitology 93, 279292.CrossRefGoogle ScholarPubMed
Zarlenga, D.S., Aschenbrenner, R.A. & Lichtenfels, J.R. (1996) Variations in microsatellite sequences provide evidence for population differences and multiple ribosomal gene repeats within Trichinella pseudospiralis. Journal of Parasitology 82, 534538.CrossRefGoogle ScholarPubMed
Zarlenga, D.S., Chute, M.B., Martin, A. & Kapel, C.M.O. (1999) A multiplex PCR for unequivocal differentiation of six encapsulated and three non-encapsulated genotypes of Trichinella. International Journal for Parasitology 29, 141149.CrossRefGoogle Scholar
Zarlenga, D.S., Rosenthal, B.M., La Rosa, G., Pozio, E. & Hoberg, E.P. (2006) Post-Miocene expansion, colonization, and host switching drove speciation among extant nematodes of the archaic genus Trichinella. Proceedings of the National Academy of Sciences USA 103, 73547359.CrossRefGoogle ScholarPubMed
Zhang, L.H., Chai, J.J., Jiao, W., Osman, Y. & McManus, D.P. (1998) Mitochondrial genomic markers confirm the presence of the camel strain (G6 genotype) of Echinococcus granulosus in north-western China. Parasitology 116, 2933.CrossRefGoogle ScholarPubMed