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Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1997 Oct;35(10):2542–2545. doi: 10.1128/jcm.35.10.2542-2545.1997

Microsatellite in the beta-tubulin gene of Toxoplasma gondii as a new genetic marker for use in direct screening of amniotic fluids.

J M Costa 1, M L Dardé 1, B Assouline 1, M Vidaud 1, S Bretagne 1
PMCID: PMC230007  PMID: 9316904

Abstract

To examine the correlation between Toxoplasma gondii genotype and congenital human toxoplasmosis, the polymorphism of the microsatellite consisting of a dinucleotide (TG) repeat in the intron of the beta-tubulin gene was investigated by PCR. Thirty-four reference strains were studied, including 7 strains virulent in mice and 27 strains avirulent in mice. The seven virulent strains had a (TG)8 microsatellite, and the avirulent strains had a (TG)7 microsatellite. This confirms the dichotomy already observed for virulent and avirulent strains. Additionally, 37 samples of amniotic fluid from infected fetuses were tested. All of them had the (TG)7 microsatellite marker. This result confirms that most of the human cases of congenital toxoplasmosis are due to strains avirulent in mice. Nevertheless, their virulence in human fetuses was obvious, as numerous abnormalities were observed on ultrasonic examination. The new genetic marker is the first one directly used for typing T. gondii isolates without any bias due to cultivation of the parasite. This microsatellite marker is not sufficient to type the strains which are avirulent in mice; however, seeking more polymorphic microsatellites should be worthwhile to obtain new genetic markers for direct screening of biological samples.

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Selected References

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  1. Asai T., Miura S., Sibley L. D., Okabayashi H., Takeuchi T. Biochemical and molecular characterization of nucleoside triphosphate hydrolase isozymes from the parasitic protozoan Toxoplasma gondii. J Biol Chem. 1995 May 12;270(19):11391–11397. doi: 10.1074/jbc.270.19.11391. [DOI] [PubMed] [Google Scholar]
  2. Bohne W., Gross U., Heesemann J. Differentiation between mouse-virulent and -avirulent strains of Toxoplasma gondii by a monoclonal antibody recognizing a 27-kilodalton antigen. J Clin Microbiol. 1993 Jun;31(6):1641–1643. doi: 10.1128/jcm.31.6.1641-1643.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bruford M. W., Wayne R. K. Microsatellites and their application to population genetic studies. Curr Opin Genet Dev. 1993 Dec;3(6):939–943. doi: 10.1016/0959-437x(93)90017-j. [DOI] [PubMed] [Google Scholar]
  4. Daffos F., Forestier F., Capella-Pavlovsky M., Thulliez P., Aufrant C., Valenti D., Cox W. L. Prenatal management of 746 pregnancies at risk for congenital toxoplasmosis. N Engl J Med. 1988 Feb 4;318(5):271–275. doi: 10.1056/NEJM198802043180502. [DOI] [PubMed] [Google Scholar]
  5. Dardé M. L. Biodiversity in Toxoplasma gondii. Curr Top Microbiol Immunol. 1996;219:27–41. doi: 10.1007/978-3-642-51014-4_3. [DOI] [PubMed] [Google Scholar]
  6. Dardé M. L., Bouteille B., Pestre-Alexandre M. Isoenzyme analysis of 35 Toxoplasma gondii isolates and the biological and epidemiological implications. J Parasitol. 1992 Oct;78(5):786–794. [PubMed] [Google Scholar]
  7. Deckert-Schlüter M., Schlüter D., Schmidt D., Schwendemann G., Wiestler O. D., Hof H. Toxoplasma encephalitis in congenic B10 and BALB mice: impact of genetic factors on the immune response. Infect Immun. 1994 Jan;62(1):221–228. doi: 10.1128/iai.62.1.221-228.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Desmonts G., Couvreur J. Congenital toxoplasmosis. A prospective study of 378 pregnancies. N Engl J Med. 1974 May 16;290(20):1110–1116. doi: 10.1056/NEJM197405162902003. [DOI] [PubMed] [Google Scholar]
  9. Dib C., Fauré S., Fizames C., Samson D., Drouot N., Vignal A., Millasseau P., Marc S., Hazan J., Seboun E. A comprehensive genetic map of the human genome based on 5,264 microsatellites. Nature. 1996 Mar 14;380(6570):152–154. doi: 10.1038/380152a0. [DOI] [PubMed] [Google Scholar]
  10. Field D., Wills C. Long, polymorphic microsatellites in simple organisms. Proc Biol Sci. 1996 Feb 22;263(1367):209–215. doi: 10.1098/rspb.1996.0033. [DOI] [PubMed] [Google Scholar]
  11. Guo Z. G., Johnson A. M. Genetic characterization of Toxoplasma gondii strains by random amplified polymorphic DNA polymerase chain reaction. Parasitology. 1995 Aug;111(Pt 2):127–132. doi: 10.1017/s0031182000064866. [DOI] [PubMed] [Google Scholar]
  12. Hohlfeld P., Daffos F., Costa J. M., Thulliez P., Forestier F., Vidaud M. Prenatal diagnosis of congenital toxoplasmosis with a polymerase-chain-reaction test on amniotic fluid. N Engl J Med. 1994 Sep 15;331(11):695–699. doi: 10.1056/NEJM199409153311102. [DOI] [PubMed] [Google Scholar]
  13. Howe D. K., Sibley L. D. Toxoplasma gondii comprises three clonal lineages: correlation of parasite genotype with human disease. J Infect Dis. 1995 Dec;172(6):1561–1566. doi: 10.1093/infdis/172.6.1561. [DOI] [PubMed] [Google Scholar]
  14. Makioka A., Ohtomo H. An increased DNA polymerase activity associated with virulence of Toxoplasma gondii. J Parasitol. 1995 Dec;81(6):1021–1022. [PubMed] [Google Scholar]
  15. Meisel R., Stachelhaus S., Mévélec M. N., Reichmann G., Dubremetz J. F., Fischer H. G. Identification of two alleles in the GRA4 locus of Toxoplasma gondii determining a differential epitope which allows discrimination of type I versus type II and III strains. Mol Biochem Parasitol. 1996 Oct 30;81(2):259–263. doi: 10.1016/0166-6851(96)02719-3. [DOI] [PubMed] [Google Scholar]
  16. Nagel S. D., Boothroyd J. C. The alpha- and beta-tubulins of Toxoplasma gondii are encoded by single copy genes containing multiple introns. Mol Biochem Parasitol. 1988 Jun;29(2-3):261–273. doi: 10.1016/0166-6851(88)90081-3. [DOI] [PubMed] [Google Scholar]
  17. Parmley S. F., Gross U., Sucharczuk A., Windeck T., Sgarlato G. D., Remington J. S. Two alleles of the gene encoding surface antigen P22 in 25 strains of Toxoplasma gondii. J Parasitol. 1994 Apr;80(2):293–301. [PubMed] [Google Scholar]
  18. Rinder H., Thomschke A., Dardé M. L., Löscher T. Specific DNA polymorphisms discriminate between virulence and non-virulence to mice in nine Toxoplasma gondii strains. Mol Biochem Parasitol. 1995 Jan;69(1):123–126. doi: 10.1016/0166-6851(94)00211-5. [DOI] [PubMed] [Google Scholar]
  19. Schlötterer C., Tautz D. Slippage synthesis of simple sequence DNA. Nucleic Acids Res. 1992 Jan 25;20(2):211–215. doi: 10.1093/nar/20.2.211. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Sibley L. D., Boothroyd J. C. Virulent strains of Toxoplasma gondii comprise a single clonal lineage. Nature. 1992 Sep 3;359(6390):82–85. doi: 10.1038/359082a0. [DOI] [PubMed] [Google Scholar]
  21. Suzuki Y., Wong S. Y., Grumet F. C., Fessel J., Montoya J. G., Zolopa A. R., Portmore A., Schumacher-Perdreau F., Schrappe M., Köppen S. Evidence for genetic regulation of susceptibility to toxoplasmic encephalitis in AIDS patients. J Infect Dis. 1996 Jan;173(1):265–268. doi: 10.1093/infdis/173.1.265. [DOI] [PubMed] [Google Scholar]

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