Hostname: page-component-cd9895bd7-7cvxr Total loading time: 0 Render date: 2024-12-26T06:57:19.178Z Has data issue: false hasContentIssue false
  • English
  • Français

Use of restriction fragment length polymorphisms (RFLPs) to distinguish between nematodes of pathogenic significance

Published online by Cambridge University Press:  06 April 2009

M. L. Cameron ,
P. Levy ,
T. Nutman ,
C. R. Vanamala ,
P. R. Narayanan  and
T. V. Rajan
M. L. Cameron
Affiliation:
Departments of Pathology and Microbiology and Immunology, Albert Einstein College of Medicine, 1300 Morris Park Ave., Bronx, New York 10461
P. Levy
Affiliation:
Departments of Pathology and Microbiology and Immunology, Albert Einstein College of Medicine, 1300 Morris Park Ave., Bronx, New York 10461
T. Nutman
Affiliation:
Departments of Pathology and Microbiology and Immunology, Albert Einstein College of Medicine, 1300 Morris Park Ave., Bronx, New York 10461
C. R. Vanamala
Affiliation:
Departments of Pathology and Microbiology and Immunology, Albert Einstein College of Medicine, 1300 Morris Park Ave., Bronx, New York 10461
P. R. Narayanan
Affiliation:
Departments of Pathology and Microbiology and Immunology, Albert Einstein College of Medicine, 1300 Morris Park Ave., Bronx, New York 10461
T. V. Rajan
Affiliation:
Departments of Pathology and Microbiology and Immunology, Albert Einstein College of Medicine, 1300 Morris Park Ave., Bronx, New York 10461
Get access Rights & Permissions [Opens in a new window]

Summary

The availability of restriction fragment length polymorphisms (RFLPs) would be useful for studying the extent of diversity among morphologically indistinguishable populations of filarial parasites. Such polymorphisms may be useful in correlating various physiological and clinical differences with parasite heterogeneity. In order to identify such RFLPs, we isolated DNA from microfilaria of 6 filarial species (Acanthocheilonema viteae, Brugia malayi, Brugia pahangi, Dirofilaria immitis, Litomosoides carinii and Setaria digitatum), digested the DNA with several restriction endonucleases, prepared Southern blots and probed with 32P-labelled DNA probes. The patterns of fragments generated using two restriction endonucleases, Mbo I and Tag I, in combination with two probes, rDNA from the free-living soil nematode Cenorhabditis elegans, and pBM103, an anonymous DNA probe from B. malayi, unequivocally distinguish between all 6 of the species. To ensure that the differences we observed between the species represent true interspecies variation rather than fortuitous individual variations we analysed DNA from several individual B. malayi and B. pahangi worms. The individual B. malayi worms demonstrated restriction profiles that were invariant, as did the individual B. pahangi worms, demonstrating that the differences we observed were true interspecies variations.

Type
Research Article
Information
Parasitology , Volume 96 , Issue 2 , April 1988 , pp. 381 - 390
Copyright
Copyright © Cambridge University Press 1988

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

REFERENCES

Ertmann, K. D., Unnasch, T. R., Greene, B. M., Albiez, E. J., Boateng, J., Denke, A. M., Ferraroni, J. J., Karam, M., Schulz-Key, H. & Williams, P. N. (1987). A DNA sequence specific for forest form Onchocerca volvulus. Nature, London 327, 415–16.Google Scholar
Files, J. C. & Hirsh, D. (1981). Ribosomal DNA of Caenorhabditis elegans. Journal of Molecular Biology 149, 223–240.CrossRefGoogle ScholarPubMed
Long, E. O. & Dawid, I. B. (1980). Repeated genes in eukaryotes. Annual Review of Biochemistry 49, 727–64.Google Scholar
McReynolds, L. A., DeSimone, S. M. & Williams, S. A. (1986). Cloning and comparison of repeated DNA sequences from the human filarial parasite Brugia malayi and the animal parasite Brugia pahangi. Proceedings of the National Academy of Sciences, USA 83, 797–801.Google Scholar
Otteson, E. A. (1984) Filariasis and tropical eosinophilia. In Tropical and Geographical Medicine (ed. Warren, K. S. and Mahmond, A. A.). New York: McGraw Hill.Google Scholar
Perler, F. B. & Karam, M. (1986). Cloning and characterization of two Onchocerca volvulus repeated DNA sequences. Molecular and Biochemical Parasitology 21, 171–8.CrossRefGoogle ScholarPubMed
Rajan, T. V., Halay, E. D., Potter, T. A., Evans, G. A., Seidman, J. G. & Margulies, D. H. (1983). H2 hemizygous mutants from a heterozygous cell line: role of mitotic recombination. EMBO Journal 2, 1537–42.CrossRefGoogle ScholarPubMed
Sanger, F., Nicklen, S. & Coulson, A. R. (1977). DNA sequencing with chain-terminating inhibitors. Proceedings of the National Acadamy of Sciences, USA 74, 5463.Google Scholar
Sim, B. K. L., Mak, J. W., Cheong, W. H., Sutanto, I., Kurniawan, L., Marwoto, H. A., Franke, E., Campell, J. R., Wirth, D. F. & Peissens, W. F. (1986). Identification of Brugia malayi in vectors with a species-specific DNA probe. American Journal of Tropical Medicine and Hygiene 35, 559–64.Google Scholar
Sim, B. K. L., Piessens, W. F. & Wirth, D. F. (1986). A DNA probe cloned in Escherichia coli for the identification of Brugia malayi. Molecular and Biochemical Parasitology 19, 117–23.Google Scholar