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

  • E. Rodríguez (a1), J. Nieto (a1), J.A. Castillo (a2) and T. Gárate (a1)

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.

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References

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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, 401407.
Boev, S.N., Britov, V.A. & Orlov, I.V. (1979) Species composition of Trichinella. Wiadomosci Parazytologyzne 25, 495503.
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, 231237.
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, 113120.
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. 405411in Tanner, C.E., Martínez Fernándex, A.R. & Bolas, Fernandex F. (Eds) Trichindlosis. Proceedings VIIICT, Alicante, Spain.
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, 8388.
Dick, T.A. (1983) Species and inrraspecific variation, pp. 3173in Campbell, W.C. (Ed.) Trichinella and trichinosis. New York.
Dupoy-Camet, J., Robert, F. & Soulé, C. (1993) RAPD in the genus Trichinella. Parasitology Today 9, 463464.
Feinberg, A.P. & Volgestein, B. (1983) A technique for radiolabelling DNA restriction endonuclease fragments to high specific activity. Analytical Biochemistry 132, 613.
Gárate, T., Albarrán, E., Bolás-Fernández, F., Martínez-Fernández, A.R. & Parkhouse, R.M. (1991) DNA polymorphisms within Spanish Trichinella isolates. Parasitology Research 77, 602605.
Heery, D.M., Gannon, F. & Powell, R. (1990) A simple method for subcloning DNA fragments from gel slices. Trends in Genetics 6, 173.
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, 772775.
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, 641646.
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, 11311140.
Lichtenfels, J.R., Pozio, E., Dick, T.A. & Zarlenga, D.S. (1994) Workshop on systematics of Trichinella. pp. 619623in Campbell, W.C., Pozio, E. & Bruschi, F. (Eds) Trichinellosis. Proceedings VIII ICT. Orvieto, Italy.
Pozio, E., La Rosa, G., Murrell, K.D. & Lichtenfels, J.R. (1992a) Taxonomic revision of the genus Trichinella. Journal of Parasitology 78, 654659.
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, 647653.
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, 568575.
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, 12471250.
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, 15211522.
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, 125137.
Sneath, P.M. & Sokal, R.R. (1973) Numerical taxonomy. San Francisco, Freeman W. H. & Co.
Soulé, C., Guillou, J.P., Dupoy-Camet, J., Vallet, C. & Pozio, E. (1993) Differentiation of Trichinella isolates by polymerase chain reaction. Parasitology Research 79, 461465.
Sulzer, A.J. (1965) Indirect fluorescent test for parasitic diseases. Preparation of a stable antigen from larvae of Trichinella spiralis. Journal of Parasitology 51, 717721.
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, 13351339.
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, 175178.
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, 222227.
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, 506513.
Welsh, J. & Mc Clelland, M. (1990) Fingerprinting in genomes using PCR with arbitrary primers. Nucleic Acids Research 18, 72137218.
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, 65316535.
Zarlenga, D.S. & Murrell, K.D. (1989) Molecular cloning of Trichinella spiralis ribosomal RNA genes: Application as genetic markers for isolate classification, pp. 3540in Tanner, C.E., Martínez Fernández, A.R. & Bolás Fernández, F. (Eds) Trichinellosis. Proceedings VII ICT. Alicante, Spain.

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

  • E. Rodríguez (a1), J. Nieto (a1), J.A. Castillo (a2) and T. Gárate (a1)

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