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Journal of Clinical Microbiology, April 2006, p. 1382-1389, Vol. 44, No. 4
0095-1137/06/$08.00+0     doi:10.1128/JCM.44.4.1382-1389.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Genotyping of Toxoplasma gondii by Multiplex PCR and Peptide-Based Serological Testing of Samples from Infants in Poland Diagnosed with Congenital Toxoplasmosis

Dorota Nowakowska,1,2 Iris Colón,3,4 Jack S. Remington,4,5 Michael Grigg,6 Elzbieta Golab,7 J. Wilczynski,2 and L. David Sibley1*

Department of Molecular Microbiology, Washington University School of Medicine, Saint Louis, Missouri 93110,1 Department of Fetal-Maternal Medicine and Gynecology, Research Institute Polish Mother's Memorial Hospital, Lodz, Poland,2 Department of Obstetrics and Gynecology, Stanford University School of Medicine, Stanford, California 94305,3 Department of Immunology and Infectious Diseases, Research Institute, Palo Alto Medical Foundation, Palo Alto, California 94301,4 Division of Infectious Diseases and Geographical Medicine, Stanford University School of Medicine, Stanford, California 94305,5 Departments of Medicine and Microbiology & Immunology, University of British Columbia, Vancouver, British Columbia V5Z 3J5, Canada,6 Department of Medical Parasitology, National Institute of Hygiene, Warsaw, Poland7

Received 2 October 2005/ Returned for modification 15 November 2005/ Accepted 23 January 2006

Toxoplasma gondii has a clonal population genetic structure with three (I, II, and III) lineages that predominate in North America and Europe. Type II strains cause most cases of symptomatic human infections in France and the United States, although few other regions have been adequately sampled. Here we determined the parasite genotype in amniotic fluid and cerebrospinal fluid samples from congenital toxoplasmosis cases in Poland. Nineteen confirmed congenital cases of toxoplasmosis were analyzed, including both severe and asymptomatic cases. The genotype of parasite strains causing congenital infection was determined by direct PCR amplification and restriction fragment length polymorphism analysis. Nested multiplex PCR analysis was used to type four independent polymorphic markers. The sensitivity of multiplex nested PCR was ≥25 parasites/ml in amniotic fluid and cerebral spinal fluid samples. Parasite DNA was successfully amplified in 9 of 19 samples (eight severely affected and one asymptomatic fetus). Only genotype II parasites were identified as the source of T. gondii infection based on restriction fragment length polymorphism analysis. Strains causing congenital infections were also typed indirectly based on detection of antibodies to strain-specific peptides. Serotyping indicated that 12 of 15 cases tested were caused by type II strains and these positives included both symptomatic and asymptomatic infections. Overall, the combined analysis indicated that 14 of the cases were caused by type II strains. Our results are consistent with the hypothesis that parasite burden is associated with severity of congenital toxoplasmosis and indicate that serological testing provides a promising method for genotypic analysis of toxoplasmosis.

* Corresponding author. Mailing address: Department of Molecular Microbiology, Washington University School of Medicine, 660 S. Euclid Ave., Saint Louis, MO 93110. Phone: (314) 362-8873. Fax: (314) 362-3203. E-mail: sibley{at}borcim.wustl.edu.

Journal of Clinical Microbiology, April 2006, p. 1382-1389, Vol. 44, No. 4
0095-1137/06/$08.00+0     doi:10.1128/JCM.44.4.1382-1389.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Sibley, L. D., Khan, A., Ajioka, J. W., Rosenthal, B. M. (2009). Genetic diversity of Toxoplasma gondii in animals and humans. Phil Trans R Soc B 364: 2749-2761 [Abstract] [Full Text]  
  • Sousa, S., Ajzenberg, D., Vilanova, M., Costa, J., Darde, M.-L. (2008). Use of GRA6-Derived Synthetic Polymorphic Peptides in an Immunoenzymatic Assay To Serotype Toxoplasma gondii in Human Serum Samples Collected from Three Continents. CVI 15: 1380-1386 [Abstract] [Full Text]  
  • Messaritakis, I., Detsika, M., Koliou, M., Sifakis, S., Antoniou, M. (2008). Prevalent Genotypes of Toxoplasma gondii in Pregnant Women and Patients from Crete and Cyprus. Am J Trop Med Hyg 79: 205-209 [Abstract] [Full Text]  


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