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Characterization of Spanish Trichinella isolates by random amplified polymorphic DNA (RAPD)

Published online by Cambridge University Press:  05 June 2009

E. Rodríguez ,
J. Nieto ,
J.A. Castillo  and
T. Gárate
E. Rodríguez
Affiliation:
Institute de Salud Carlos III, Centro Nacional de Microbiología, Sevicio de Parasitología, 28220 Majadahonda, Spain
J. Nieto
Affiliation:
Institute de Salud Carlos III, Centro Nacional de Microbiología, Sevicio de Parasitología, 28220 Majadahonda, Spain
J.A. Castillo
Affiliation:
Facultad de Veterinaria, Unidad de Parasitología y Enfermedades Parasitarias, Departamento de Patología Animal, Universidad de Zaragoza, Spain
T. Gárate*
Affiliation:
Institute de Salud Carlos III, Centro Nacional de Microbiología, Sevicio de Parasitología, 28220 Majadahonda, Spain
*
*Author for correspondence.
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Abstract

The random amplified polymorphic DNA (RAPD) assay was used to find molecular markers able to distinguish Trichinella spiralis from T. britovi, the two recognized Spanish Trichinella species. Fourteen Spanish Trichinella isolates, as well as reference Trichinella isolates representing the five species T. spiralis (T1), T. nativa (T2), T. britovi (T3), T. pseudospiralis (T4) and T. nelsoni (T7) and the three other taxa Trichinella T5, Trichinella T6 and Trichinella T8 of the genus, were characterized by RAPD using both purified and crude DNAs from infective muscle larvae (ML) and seven arbitrary primers. Three primers yielded diagnostic RAPD markers for the Spanish T. spiralis and T. britovi isolates as well as for the Trichinella reference isolates analysed, and in the case of crude DNAs the results were obtained in few hours. In addition, the species-specificity of the diagnostic RAPD markers from Spanish Trichinella isolates was studied by cross-hybridization assays. These assays confirmed that the selected diagnostic DNA fragments were not species-specific, but showed potential differences in the copy number among the examined Trichinella genetic clusters.

Type
Research Papers
Information
Journal of Helminthology , Volume 70 , Issue 4 , December 1996 , pp. 335 - 343
Copyright
Copyright © Cambridge University Press 1996

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References

Bandi, C., La, Rosa G., Bardin, M.G., Damiani, G., Comincini, S., Tasciotti, L. & Pozio, E. (1995) Random amplified polymorphic DNA fingerprints of the eight taxa of Trichinella and their comparision with allozyme analysis. Parasitology 110, 401–407.CrossRefGoogle Scholar
Boev, S.N., Britov, V.A. & Orlov, I.V. (1979) Species composition of Trichinella. Wiadomosci Parazytologyzne 25, 495–503.Google Scholar
Bolás, E., Albarrán, E., Gárate, T., Parkhouse, R.M.E. & Martínez, A.R. (1992) A longitudinal study of porcine serological responses to experimental infections with Tl and T3 Spanish Trichinella isolates. Journal ofHelminthology 66, 231–237.CrossRefGoogle Scholar
Chambers, A.E., Almond, N.M., Knight, M., Simpson, A.J.G. & Parkhouse, R.M.E. (1986) Repetitive DNA as tool for the identification and comparision of nematode variants: application to Trichinella isolates. Molecular and Biochemical Parasitology 21, 113–120.CrossRefGoogle Scholar
Del Real, G., Germain, J.L., Rodriguez, E., Sahagún, B., Sainz-Pardo, D. & Ramos, J. (1989) Sero-epidemiological studies on a human population submitted to high risk of Trichinella infection, pp. 405–411in Tanner, C.E., Martínez Fernándex, A.R. & Bolas, Fernandex F. (Eds) Trichindlosis. Proceedings VIIICT, Alicante, Spain.Google Scholar
Dias-Neto, E., Pereira de Souza, C., Rollinson, D., Katz, N., Pena, S.D.J. & Simpson, A.J.G. (1993) The random amplification of polymorphic DNA allows the identification of strains and species of schistosome. Molecular and Biochemical Parasitology 57, 83–88.CrossRefGoogle ScholarPubMed
Dick, T.A. (1983) Species and inrraspecific variation, pp. 31–73in Campbell, W.C. (Ed.) Trichinella and trichinosis. New York.CrossRefGoogle Scholar
Dupoy-Camet, J., Robert, F. & Soulé, C. (1993) RAPD in the genus Trichinella. Parasitology Today 9, 463–464.CrossRefGoogle Scholar
Feinberg, A.P. & Volgestein, B. (1983) A technique for radiolabelling DNA restriction endonuclease fragments to high specific activity. Analytical Biochemistry 132, 6–13.CrossRefGoogle ScholarPubMed
Gárate, T., Albarrán, E., Bolás-Fernández, F., Martínez-Fernández, A.R. & Parkhouse, R.M. (1991) DNA polymorphisms within Spanish Trichinella isolates. Parasitology Research 77, 602–605.CrossRefGoogle ScholarPubMed
Heery, D.M., Gannon, F. & Powell, R. (1990) A simple method for subcloning DNA fragments from gel slices. Trends in Genetics 6, 173.Google ScholarPubMed
Klassen, G.R., Thiessen, J.P. & Dick, T.A. (1986) Restriction endonuclease analysis of repetitive sequences in the Trichinella genome: Three strain-specific patterns. Journal of Parasitology 72, 772–775.CrossRefGoogle ScholarPubMed
La, Rosa G., Pozio, E., Rossi, P. & Murrell, K.D. (1992) Allozyme analysis of Trichinella isolates from various host species and geographical regions. Journal of Parasitology 78, 641–646.Google Scholar
Lichtenfels, J.R., Murrell, K.D. & Pilitt, P.A. (1983) Comparison of three subspecies of Trichinella spiralis by scanning electron microscopy. Journal of Parasitology 69, 1131–1140.CrossRefGoogle ScholarPubMed
Lichtenfels, J.R., Pozio, E., Dick, T.A. & Zarlenga, D.S. (1994) Workshop on systematics of Trichinella. pp. 619–623in Campbell, W.C., Pozio, E. & Bruschi, F. (Eds) Trichinellosis. Proceedings VIII ICT. Orvieto, Italy.Google Scholar
Pozio, E., La Rosa, G., Murrell, K.D. & Lichtenfels, J.R. (1992a) Taxonomic revision of the genus Trichinella. Journal of Parasitology 78, 654–659.CrossRefGoogle ScholarPubMed
Pozio, E., La Rosa, G., Rossi, P. & Murrell, K.D. (1992b) Biological characterization of Trichinella isolates from various host species and geographical regions. Journal of Parasitology 78, 647–653.CrossRefGoogle ScholarPubMed
Pozio, E., Varese, P.M., Gomez-Morales, M.A., Croppo, G.P., Pelliccia, D. & Bruschi, F. (1993) Comparison of human trichinellosis caused by Trichinella spiralis and by Trichinella britovi. American Journal of Tropical Medicine and Hygiene 48, 568–575.CrossRefGoogle ScholarPubMed
Pozio, E., Bandi, C., La Rosa, G., Jarvis, T., Miller, I. & Kapel, C.M. (1995) Concurrent infection with sibling Trichinella species in a natural host. International Journal for Parasitology 25, 1247–1250.CrossRefGoogle Scholar
Rodríguez, E., Nieto, J., Rodríguez, M. & Gárate, T. (1995) Use of Random Amplified Polymorphic DNA for detection of Trichinella britovi outbreaks in Spain. Clinical Infectious Diseases 21, 1521–1522.CrossRefGoogle ScholarPubMed
Simpson, A.J., Sher, A. & Me Cutchan, T.F. (1982) The genome of Schistosoma mansoni: isolation, size, bases and repetitive sequences. Molecular and Biodiemical Parasitology 6, 125–137.CrossRefGoogle ScholarPubMed
Sneath, P.M. & Sokal, R.R. (1973) Numerical taxonomy. San Francisco, Freeman W. H. & Co.Google Scholar
Soulé, C., Guillou, J.P., Dupoy-Camet, J., Vallet, C. & Pozio, E. (1993) Differentiation of Trichinella isolates by polymerase chain reaction. Parasitology Research 79, 461–465.CrossRefGoogle ScholarPubMed
Sulzer, A.J. (1965) Indirect fluorescent test for parasitic diseases. Preparation of a stable antigen from larvae of Trichinella spiralis. Journal of Parasitology 51, 717–721.CrossRefGoogle Scholar
Tibayrenc, M., Neubauer, K., Bernabe, C., Guerrini, F., Skarecky, D. & Ayala, F. (1993) Genetic characterization of six parasitic protozoa: Parity between random primer DNA typing and multilocus enzyme electrophoresis. Proceedings of the National Academy of Sciences of the United States of America 90, 1335–1339.CrossRefGoogle ScholarPubMed
Tighe, P.J., Goyal, P.K., Willson, 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, 175–178.CrossRefGoogle ScholarPubMed
Vahidi, H., Curran, J., Nelson, D.W., Webster, J.M., McClure, M.A. & Honda, B.M. (1988) Unusual sequences, homologous to 5S RNA, in ribosomal DNA repeats of the nematodeMe/ofdogyne arenaria. Journal of Molecular Evolution 27, 222–227.CrossRefGoogle ScholarPubMed
Walsh, P.S., Metzger, D.A. & Higuchi, R. (1991) Chelex-100 as a medium for simple extraction of DNA for PCR based typing from forensic material. Biotechniques 10, 506–513.Google ScholarPubMed
Welsh, J. & Mc Clelland, M. (1990) Fingerprinting in genomes using PCR with arbitrary primers. Nucleic Acids Research 18, 7213–7218.CrossRefGoogle ScholarPubMed
Williams, J.G.K., Kubelik, A., Livak, K.J., Rifalski, J.A. & Tingey, S.V. (1990) DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Research 18, 6531–6535.CrossRefGoogle ScholarPubMed
Zarlenga, D.S. & Murrell, K.D. (1989) Molecular cloning of Trichinella spiralis ribosomal RNA genes: Application as genetic markers for isolate classification, pp. 35–40in Tanner, C.E., Martínez Fernández, A.R. & Bolás Fernández, F. (Eds) Trichinellosis. Proceedings VII ICT. Alicante, Spain.Google Scholar