Skip to main content Accessibility help
×
×
Home

Moving towards an integrated approach to molecular detection and identification of Toxoplasma gondii

  • C. SU (a1), E. K. SHWAB (a1), P. ZHOU (a2), X. Q. ZHU (a2) and J. P. DUBEY (a3)...
Summary

The development of simple, sensitive and rapid methods for the detection and identification of Toxoplasma gondii is important for the diagnosis and epidemiological studies of the zoonotic disease toxoplasmosis. In the past 2 decades, molecular methods based on a variety of genetic markers have been developed, each with its advantages and limitations. The application of these methods has generated invaluable information to enhance our understanding of the epidemiology, population genetics and phylogeny of T. gondii. However, since most studies focused solely on the detection but not genetic characterization of T. gondii, the information obtained was limited. In this review, we discuss some widely used molecular methods and propose an integrated approach for the detection and identification of T. gondii, in order to generate maximum information for epidemiological, population and phylogenetic studies of this key pathogen.

Copyright
Corresponding author
*Corresponding author: Department of Microbiology, the University of Tennessee, Knoxville, TN 37966, USA. Tel: +865 974 4015. Fax: +865 974 4007. E-mail: csu1@utk.edu
References
Hide All
Ajzenberg, D., Banuls, A. L., Su, C., Dumetre, A., Demar, M., Carme, B. and Darde, M. L. (2004). Genetic diversity, clonality and sexuality in Toxoplasma gondii. International Journal for Parasitology 34, 11851196.
Ajzenberg, D., Banuls, A. L., Tibayrenc, M. and Darde, M. L. (2002 a). Microsatellite analysis of Toxoplasma gondii shows considerable polymorphism structured into two main clonal groups. International Journal for Parasitology 32, 2738.
Ajzenberg, D., Cogne, N., Paris, L., Bessieres, M. H., Thulliez, P., Filisetti, D., Pelloux, H., Marty, P. and Darde, M. L. (2002 b). Genotype of 86 Toxoplasma gondii isolates associated with human congenital toxoplasmosis, and correlation with clinical findings. Journal of Infectious Diseases 186, 684689.
Ajzenberg, D., Dumetre, A. and Darde, M.-L. (2005). Multiplex PCR for typing strains of Toxoplasma gondii. Journal of Clinical Microbiology 43, 19401943.
Bastien, P., Jumas-Bilak, E., Varlet-Marie, E. and Marty, P. (2007). Three years of multi-laboratory external quality control for the molecular detection of Toxoplasma gondii in amniotic fluid in France. Clinical Microbiology and Infection 13, 430433.
Bell, A. and Ranford-Cartwright, L. (2002). Real-time quantitative PCR in parasitology. Trends in Parasitology 18, 337342.
Bessières, M. H., Berrebi, A., Cassaing, S., Fillaux, J., Cambus, J. P., Berry, A., Assouline, C., Ayoubi, J. M. and Magnaval, J. F. (2009). Diagnosis of congenital toxoplasmosis: prenatal and neonatal evaluation of methods used in Toulouse University Hospital and incidence of congenital toxoplasmosis. Memorias do Instituto Oswaldo Cruz 104, 389392.
Blackston, C. R., Dubey, J. P., Dotson, E., Su, C., Thulliez, P., Sibley, D. and Lehmann, T. (2001). High-resolution typing of Toxoplasma gondii using microsatellite loci. Journal of Parasitology 87, 14721475.
Bossi, P. and Bricaire, F. (2004). Severe acute disseminated toxoplasmosis. The Lancet 364, 579.
Bretagne, S., Costa, J. M., Fleury-Feith, J., Poron, F., Dubreuil-Lemaire, M. L. and Vidaud, M. (1995). Quantitative competitive PCR with bronchoalveolar lavage fluid for diagnosis of toxoplasmosis in AIDS patients. Journal of Clinical Microbiology 33, 16621664.
Burg, J. L., Grover, C. M., Pouletty, P. and Boothroyd, J. C. (1989). Direct and sensitive detection of a pathogenic protozoan Toxoplasma gondii, by polymerase chain reaction. Journal of Clinical Microbiology 27, 17871792.
Calderaro, A., Piccolo, G., Gorrini, C., Peruzzi, S., Zerbini, L., Bommezzadri, S., Dettori, G. and Chezzi, C. (2006). Comparison between two real-time PCR assays and a nested-PCR for the detection of Toxoplasma gondii. Acta Bio Medica 77, 7580.
Contini, C., Cultrera, R., Seraceni, S., Segala, D., Romani, R., Fainardi, E., Cinque, P., Lazzarin, A. and Delia, S. (2002). The role of stage-specific oligonucleotide primers in providing effective laboratory support for the molecular diagnosis of reactivated Toxoplasma gondii encephalitis in patients with AIDS. Journal of Medical Microbiology 51, 879890.
Contini, C., Seraceni, S., Cultrera, R., Incorvaia, C., Sebastiani, A. and Picot, S. (2005). Evaluation of a Real-time PCR-based assay using the lightcycler system for detection of Toxoplasma gondii bradyzoite genes in blood specimens from patients with toxoplasmic retinochoroiditis. International Journal for Parasitology 35, 275283.
Costa, J. M., Pautas, C., Ernault, P., Foulet, F., Cordonnier, C. and Bretagne, S. (2000). Real-time PCR for diagnosis and follow-up of Toxoplasma reactivation after allogeneic stem cell transplantation using fluorescence resonance energy transfer hybridization probes. Journal of Clinical Microbiology 38, 29292932.
Darde, M. L., Bouteille, B. and Pestre-Alexandre, M. (1992). Isoenzyme analysis of 35 Toxoplasma gondii isolates and the biological and epidemiological implications. Journal of Parasitology 78, 786794.
Dubey, J. P. (2004). Toxoplasmosis – a waterborne zoonosis. Veterinary Parasitology 126, 5772.
Dubey, J. P., Applewhaite, L., Sundar, N., Velmurugan, G. V., Bandini, L. A., Kwok, O. C., Hill, R. and Su, C. (2007 g). Molecular and biological characterization of Toxoplasma gondii isolates from free-range chickens from Guyana, South America, identified several unique and common parasite genotypes. Parasitology 134, 15591565.
Dubey, J. P. and Beattie, C. P. (1988). Toxoplasmosis of Animals and Man. CRC Press, Boca Raton, FL, USA.
Dubey, J. P., Cortés-Vecino, J. A., Vargas-Duarte, J. J., Sundar, N., Velmurugan, G. V., Bandini, L. M., Polo, L. J., Zambrano, L., Mora, L. E., Kwok, O. C. H., Smith, T. and Su, C. (2007 a). Prevalence of Toxoplasma gondii in dogs from Colombia, South America and genetic characterization of T. gondii isolates. Veterinary Parasitology 145, 4550.
Dubey, J. P., Hill, D. E., Sundar, N., Velmurugan, G. V., Bandini, L. A., Kwok, O. C., Pierce, V., Kelly, K., Dulin, M., Thulliez, P., Iwueke, C. and Su, C. (2008 a). Endemic toxoplasmosis in pigs on a farm in Maryland: isolation and genetic characterization of Toxoplasma gondii. Journal of Parasitology 94, 3641.
Dubey, J. P., Huong, L. T. T., Sundar, N. and Su, C. (2007 b). Genetic characterization of Toxoplasma gondii isolates in dogs from Vietnam suggests their South American origin. Veterinary Parasitology 146, 347351.
Dubey, J. P., Lopez-Torres, H. Y., Sundar, N., Velmurugan, G. V., Ajzenberg, D., Kwok, O. C. H., Hill, R., Darde, M. L. and Su, C. (2007 c). Mouse-virulent Toxoplasma gondii isolated from feral cats on Mona Island, Puerto Rico. Journal of Parasitology 93, 13651369.
Dubey, J. P., Quirk, T., Pittt, J. A., Sundar, N., Velmurugan, G. V., Kwok, O. C., Hill, R. and Su, C. (2008 b). Isolation and genetic characterization of Toxoplasma gondii from raccoons (Procyon lotor), cats (Felis domesticus), striped skunk (Mephitis mephitis), black bear (Ursus americanus), and cougar (Puma concolor) from Canada. Journal of Parasitology 94, 4245.
Dubey, J. P., Rajapakse, R. P. V. J., Wijesundera, R. R. M. K. K., Sundar, N., Velmurugan, G. V., Kwok, O. C. H. and Su, C. (2007 d). Prevalence of Toxoplasma gondii in dogs from Sri Lanka and genetic characterization of the parasite isolates. Veterinary Parasitology 146, 341346.
Dubey, J. P. and Su, C. (2009). Population biology of Toxoplasma gondii: what's out and where did they come from. Memorias do Instituto Oswaldo Cruz 104, 190195.
Dubey, J. P., Sundar, N., Gennari, S. M., Minervino, A. H. H., Farias, N. A. D. R., Ruas, J. L., Dos Santos, T. R. B., Cavalcante, G. T., Kwok, O. C. H. and Su, C. (2007 e). Biologic and genetic comparison of Toxoplasma gondii isolates in free-range chickens from the northern Para state and the southern state Rio Grande do Sul, Brazil revealed highly diverse and distinct parasite populations. Veterinary Parasitology 143, 182188.
Dubey, J. P., Sundar, N., Hill, D., Velmurugan, G. V., Bandini, L. A., Kwok, O. C. H., Majumdar, D. and Su, C. (2008 c). High prevalence and abundant atypical genotypes of Toxoplasma gondii isolated from lambs destined for human consumption in the USA. International Journal for Parasitology 38, 999–1006.
Dubey, J. P., Sundar, N., Nolden, C. A., Samuel, M. D., Velmurugan, G. V., Bandini, L. A., Kwok, O. C. H., Bodenstein, B. and Su, C. (2007 f). Characterization of Toxoplasma gondii from raccoons (Procyon lotor), coyotes (Canis latrans), and striped skunks (Mephitis mephitis) in Wisconsin identified several atypical genotypes. Journal of Parasitology 93, 15241527.
Dubey, J. P., Velmurugan, G. V., Chockalingam, A., Pena, H. F. J., De Oliveira, L. N., Leifer, C. A., Gennari, S. M., Bahia Oliveira, L. M. G. and Su, C. (2008 d). Genetic diversity of Toxoplasma gondii isolates from chickens from Brazil. Veterinary Parasitology 157, 299305.
Dubey, J. P., Velmurugan, G., Morales, J. A., Arguedas, R. and Su, C. (2009). Isolation of Toxoplasma gondii from the keel-billed toucan (Ramphastos sulfuratus) from Costa Rica. Journal of Parasitology 95, 467468.
Dubey, J. P., Velmurugan, G. V., Ulrich, V., Gill, J., Carstensen, M., Sundar, N., Kwok, O. C. H., Thulliez, P., Majumdar, D. and Su, C. (2008 e). Transplacental toxoplasmosis in naturally-infected white-tailed deer: Isolation and genetic characterisation of Toxoplasma gondii from foetuses of different gestational ages. International Journal for Parasitology 38, 10571063.
Edvinsson, B., Lappalainen, M. and Evengard, B. (2006). Real-time PCR targeting a 529-bp repeat element for diagnosis of toxoplasmosis. Clinical Microbiology and Infection 12, 131136.
Fazaeli, A., Carter, P. E., Darde, M. L. and Pennington, T. H. (2000). Molecular typing of Toxoplasma gondii strains by GRA6 gene sequence analysis. International Journal for Parasitology 30, 637642.
Ferreira, A. D. M., Vitor, R. W., Gazzinelli, R. T. and Melo, M. N. (2006). Genetic analysis of natural recombinant Brazilian Toxoplasma gondii strains by multilocus PCR-RFLP. Infection, Genetics and Evolution 6, 2231.
Ferreira, I. M., Vidal, J. E., Costa-Silva, T. A., Meira, C. S., Hiramoto, R. M., Penalva De Oliveira, A. C. and Pereira-Chioccola, V. L. (2008). Toxoplasma gondii: genotyping of strains from Brazilian AIDS patients with cerebral toxoplasmosis by multilocus PCR-RFLP markers. Experimental Parasitology 118, 221227.
Goldstein, D. B. and Schlotterer, C. (1999). Microsatellites: Evolution and Applications, Oxford University Press, Oxford, UK.
Grigg, M. E., Ganatra, J., Boothroyd, J. C. and Margolis, T. P. (2001). Unusual abundance of atypical strains associated with human ocular toxoplasmosis. Journal of Infectious Diseases 184, 633639.
Homan, W. L., Vercammen, M., De Braekeleer, J. and Verschueren, H. (2000). Identification of a 200- to 300-fold repetitive 529 bp DNA fragment in Toxoplasma gondii, and its use for diagnostic and quantitative PCR. International Journal for Parasitology 30, 6975.
Howe, D. K., Honore, S., Derouin, F. and Sibley, L. D. (1997). Determination of genotypes of Toxoplasma gondii strains isolated from patients with toxoplasmosis. Journal of Clinical Microbiology 35, 14111414.
Howe, D. K. and Sibley, L. D. (1995). Toxoplasma gondii comprises three clonal lineages: correlation of parasite genotype with human disease. Journal of Infectious Diseases 172, 15611566.
Hurtado, A., Aduriz, G., Moreno, B., Barandika, J. and Garcia-Perez, A. L. (2001). Single tube nested PCR for the detection of Toxoplasma gondii in fetal tissues from naturally aborted ewes. Veterinary Parasitology 102, 1727.
Jauregui, L. H., Higgins, J., Zarlenga, D., Dubey, J. P. and Lunney, J. K. (2001). Development of a real-time PCR assay for detection of Toxoplasma gondii in pig and mouse tissues. Journal of Clinical Microbiology 39, 20652071.
Kasper, D. C., Sadeghi, K., Prusa, A.-R., Reischer, G. H., Kratochwill, K., Förster-Waldl, E., Gerstl, N., Hayde, M., Pollak, A. and Herkner, K. R. (2009). Quantitative real-time polymerase chain reaction for the accurate detection of Toxoplasma gondii in amniotic fluid. Diagnostic Microbiology and Infectious Disease 63, 1015.
Khalifa, K. E. S., Roth, A., Roth, B., Arasteh, K. N. and Janitschke, K. (1994). Value of PCR for evaluating occurrence of parasitemia in immunocompromised patients with cerebral and extracerebral toxoplasmosis. Journal of Clinical Microbiology 32, 28132819.
Khan, A., Fux, B., Su, C., Dubey, J. P., Darde, M. L., Ajioka, J. W., Rosenthal, B. M. and Sibley, L. D. (2007). Recent transcontinental sweep of Toxoplasma gondii driven by a single monomorphic chromosome. Proceedings of the National Academy of Sciences, USA 104, 1487214877.
Khan, A., Jordan, C., Muccioli, C., Vallochi, A. L., Rizzo, L. V., Belfort, R. Jr., Vitor, R. W. A., Silveira, C. and Sibley, L. D. (2006). Genetic divergence of Toxoplasma gondii strains associated with ocular Toxoplasmosis, Brazil. Emerging and Infectious Diseases 12, 942949.
Khan, A., Su, C., German, M., Storch, G. A., Clifford, D. B. and Sibley, L. D. (2005 a). Genotyping of Toxoplasma gondii Strains from immunocompromised patients reveals high prevalence of type I strains. Journal of Clinical Microbiology 43, 58815887.
Khan, A., Taylor, S., Su, C., Mackey, A. J., Boyle, J., Cole, R., Glover, D., Tang, K., Paulsen, I. T., Berriman, M., Boothroyd, J. C., Pfefferkorn, E. R., Dubey, J. P., Ajioka, J. W., Roos, D. S., Wootton, J. C. and Sibley, L. D. (2005 b). Composite genome map and recombination parameters derived from three archetypal lineages of Toxoplasma gondii. Nucleic Acids Research 33, 29802992.
Kupferschmidt, O., Kruger, D., Held, T. K., Ellerbrok, H., Siegert, W. and Janitschke, K. (2001). Quantitative detection of Toxoplasma gondii DNA in human body fluids by TaqMan polymerase chain reaction. Clinical Microbiology and Infection 7, 120124.
Lehmann, T., Graham, D. H., Dahl, E. R., Bahia-Oliveira, L. M., Gennari, S. M. and Dubey, J. P. (2004). Variation in the structure of Toxoplasma gondii and the roles of selfing, drift, and epistatic selection in maintaining linkage disequilibria. Infection, Genetics and Evolution 4, 107114.
Lehmann, T., Marcet, P. L., Graham, D. H., Dahl, E. R. and Dubey, J. P. (2006). Globalization and the population structure of Toxoplasma gondii. Proceedings of the National Academy of Sciences, USA 103, 1142311428.
Liesenfeld, O., Roth, A., Weinke, T., Foss, H. D. and Hahn, H. (1994). A case of disseminated toxoplasmosis-value of PCR for the diagnosis. Journal of Infection 29, 133138.
Lin, M. H., Chen, T. C., Kuo, T., Tseng, C. C. and Tseng, C. P. (2000). Real-time PCR for quantitative detection of Toxoplasma gondii. Journal of Clinical Microbiology 38, 41214125.
Montoya, J. G. and Liesenfeld, O. (2004). Toxoplasmosis. The Lancet 363, 19651976.
Okay, T. S., Yamamoto, L., Oliveira, L. C., Manuli, E. R., Andrade Junior, H. F. D. and Del Negro, G. M. B. (2009). Significant performance variation among PCR systems in diagnosing congenital toxoplasmosis in São Paulo, Brazil: analysis of 467 amniotic fluid samples. Clinics 64, 171176.
Pelloux, H., Guy, D., Angelici, M. C., Aspock, H., Bessiéres, M., Blatz, R., Del Pezzo, M., Girault, V., Gratzl, R., Holberg-Petersen, M., Johnson, J., Kruger, D., Lappalainen, M., Naessens, A. and Olsson, M. (1998). A second European collaborative study on polymerase chain reaction for Toxoplasma gondii, involving 15 teams. FEMS Microbiology Letters 165, 231237.
Pena, H. F. J., Gennari, S. M., Dubey, J. P. and Su, C. (2008). Population structure and mouse-virulence of Toxoplasma gondii in Brazil. International Journal for Parasitology 38, 561569.
Prestrud, K. W., Åsbakk, K., Mørk, T., Fuglei, E., Tryland, M. and Su, C. (2008). Direct high-resolution genotyping of Toxoplasma gondii in arctic foxes (Vulpes lagopus) in the remote arctic Svalbard archipelago reveals widespread clonal type II lineage. Veterinary Parasitology 158, 121128.
Reischl, U., Bretagne, S., Kruger, D., Ernault, P. and Costa, J. M. (2003). Comparison of two DNA targets for the diagnosis of Toxoplasmosis by real-time PCR using fluorescence resonance energy transfer hybridization probes. BMC Infectious Diseases 3, 7.
Schoondermark-Van De Ven, E., Melchers, W., Camps, W., Eskes, T., Meuwissen, J. and Galama, J. (1994). Effectiveness of spiramycin for treatment of congenital Toxoplasma gondii infection in Rhesus monkeys. Antimicrobial Agents and Chemotherapy 38, 19301936.
Sibley, L. D. and Boothroyd, J. C. (1992). Virulent strains of Toxoplasma gondii comprise a single clonal lineage. Nature, London 359, 8285.
Su, C. and Dubey, J. P. (2009). Toxoplasma. In Molecular Detection of Foodborne Pathogens (ed. Liu, D.), pp. 741753. CRC Press, Boca Raton, FL, USA.
Su, C., Zhang, X. and Dubey, J. P. (2006). Genotyping of Toxoplasma gondii by multilocus PCR-RFLP markers: A high resolution and simple method for identification of parasites. International Journal for Parasitology 36, 841848.
Sundar, N., Cole, R. A., Thomas, N. J., Majumdar, D., Dubey, J. P. and Su, C. (2008). Genetic diversity among sea otter isolates of Toxoplasma gondii. Veterinary Parasitology 151, 125132.
Switaj, K., Master, A., Skrzypczak, M. and Zaborowski, P. (2005). Recent trends in molecular diagnostics for Toxoplasma gondii infections. Clinical Microbiology and Infection 11, 170176.
Tenter, A. M., Heckeroth, A. R. and Weiss, L. M. (2000). Toxoplasma gondii: from animals to humans. International Journal for Parasitology 30, 12171258.
Velmurugan, G. V., Dubey, J. P. and Su, C. (2008). Genotyping studies of Toxoplasma gondii isolates from Africa revealed that the archetypal clonal lineages predominate as in North America and Europe. Veterinary Parasitology 155, 314318.
Recommend this journal

Email your librarian or administrator to recommend adding this journal to your organisation's collection.

Parasitology
  • ISSN: 0031-1820
  • EISSN: 1469-8161
  • URL: /core/journals/parasitology
Please enter your name
Please enter a valid email address
Who would you like to send this to? *
×

Keywords

Metrics

Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed