Journal
JOURNAL OF CLINICAL MICROBIOLOGY
Volume 43, Issue 12, Pages 5881-5887Publisher
AMER SOC MICROBIOLOGY
DOI: 10.1128/JCM.43.12.5881-5887.2005
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Funding
- NIAID NIH HHS [AI 36629, R01 AI036629, U01 AI038858, R01 AI059176, AI 38858] Funding Source: Medline
- NIMH NIH HHS [MH 22005] Funding Source: Medline
- NINDS NIH HHS [NS 32228, R01 NS032228, U01 NS032228] Funding Source: Medline
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Toxoplasma gondii is an important food- and waterborne opportunistic pathogen that causes severe disease in immunocom promised patients. T. gondii has an unusual clonal population structure consisting of three widespread lineages known as I, II, and III. To establish the genotypes of strains of T. gondii associated with human toxoplasmosis, we have developed a set of four highly sensitive and polymorphic nested PCR markers. Multiplex nested PCR analysis was used to genotype parasites in cerebral spinal fluid samples from 8 of 10 human immunodeficiency virus-positive patients. Remarkably, a majority of these patients had infections with type I strains or strains containing type I alleles, despite the fact that this lineage is normally uncommon in humans and animals. Multiplex analysis of these four unlinked makers was able to distinguish all three common genotypes and also detected two strains with mixed genotypes. Further analysis based on sequencing of a polymorphic intron revealed that one of these recombinant strains was an exotic lineage distinct from the archetypal clonal lineages. The multiplex nested PCR analysis described here will be useful for analyzing the contribution of parasite genotype to toxoplasmosis.
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