Genetic variability of Brazilian Toxoplasma gondii strains detected by random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) and simple sequence repeat anchored-PCR (SSR-PCR)

Ferreira, Adriana de Melo; Vítor, Ricardo Wagner Almeida; Carneiro, Ana Carolina Aguiar Vasconcelos; Brandão, G.P.; Melo, Maria Norma

doi:10.1016/j.meegid.2004.03.002

Cited by 47 publications

(22 citation statements)

References 60 publications

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“…Six T. gondii strains isolated in Minas Gerais, Brazil and previously genotyped by PCR-RFLP assay of the SAG2 gene were used in this study: SAF, EGS and D4 (type I) and D7, CH1 and CH3 (type III), in addition to the reference RH strain (type I), were used (Ferreira et al, 2004). Type II strains were not used in this study as, so far, no strain of this genotype has been isolated in Brazil.…”

Section: T Gondii Isolatesmentioning

confidence: 99%

Efficacy of atovaquone and sulfadiazine in the treatment of mice infected withToxoplasma gondiistrains isolated in Brazil

Alves

Vítor

2005

Parasite

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Summary :The efficacy of atovaquone and sulfadiazine was examined alone or in combination for the treatment of mice infected with six Brazilian Toxoplasma gondii strains previously genotyped using the PCR-RFLP assays of the SAG2 gene, in addition to RH strain. Swiss mice were infected intraperitoneally with 10 2 tachyzoites from each strain of T. gondii and treated with 6.25, 12.5, 25 and 50 mg/Kg/day of atovaquone or 40, 80, 160 and 320 mg/Kg/day of sulfadiazine. In a second experiment, mice were treated with the association of previously determined doses of each drug. Treatment started 48 hours post-infection, and lasted 10 days. The susceptibility of T. gondii to atovaquone and to sulfadiazine was different according to the parasite strain. It was observed strains that are susceptible to atovaquone, and strains that are resistant to it. Type I strains were more susceptible to the activity of sulfadiazine and more resistant to atovaquone. Yet type III strains were susceptible to atovaquone and to sulfadiazine. Association of atovaquone and sulfadiazine presented a synergic effect in the treatment of mice infected with RH type I strain and an additive effect in the treatment of mice infected with one type I strain and with two type III strains. Résumé

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Section: T Gondii Isolatesmentioning

confidence: 99%

Efficacy of atovaquone and sulfadiazine in the treatment of mice infected withToxoplasma gondiistrains isolated in Brazil

Alves

Vítor

2005

Parasite

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“…Ferreira et al (2006), using a multilocus PCR-RFLP, demonstrated that Brazilian strains present recombinant genotypes with typical alleles of the strains types I, II, and III in the majority of the loci. The RAPD-PCR and SSR-PCR analysis of these same strains placed them into two different clusters correlated with virulence for BALB/c mice (Ferreira et al, 2004). The aim of the current study was to obtain T. gondii isolates directly from tissues of pigs, dogs and chickens in the State of Minas Gerais, Brazil, and to determine their virulence in BALB/c mice.…”

Section: Introductionmentioning

confidence: 99%

Characterization ofToxoplasma gondiifrom domestic animals from Minas Gerais, Brazil

Brandão

Ferreira²,

Melo³

et al. 2006

Parasite

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Summary :In an attempt to isolate and characterize Toxoplasma gondii from the State of Minas Gerais, Brazil, musculature samples from 72 pigs, 25 dogs, 28 free-range chickens and 50 chickens produced in industrialized farms were collected. Antibodies to T. gondii have not been detected in pigs, but were found in nine (40.9 %) out of 22 dogs, and in 15 (53.6 %) of 28 free range chickens. T. gondii was not isolated from pigs and industrialized chickens, but from eight dogs and 11 free range chickens. In order to determine T. gondii virulence, female BALB/c mice were inoculated with 10 3 , 10 2 , 10 1 and 10 0 tachyzoites of the 19 isolates. The strains RH (virulent) and ME49 (non-virulent) were used as references. Isolates were divided into three groups according to the virulence phenotype: five isolates were classified into virulent in mice, one into non-virulent and 13 into intermediate virulent. Nested-PCR of T. gondii SAG2 locus amplified DNA from 21 out of 22 DNA samples directly extracted from heart of free range chickens. These samples were genotyped through a PCR-RFLP assay. Seventeen (80.9 %) were classified into type I; one (4.8 %) into type III and three (14.3 %) into type I or II. Résumé

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“…They are highly polymorphic, the MS length varies depending the numbers of STR. The tandem repetitions in T. gondii are often simple, with only repetitions of two nucleotides, which occur from 2 to 20 times [62,63]. Around 15 markers of MS have been used, including (tub2, W35, TgM-A, B18, B17; M33, IV.1, XI.1, M48, M102, N60, N82, AA, N61, N83) to determine the genotype of T. gondii in diferent laboratories [61,62].…”

Section: Polymerase Chain Reaction-restriction Fragment Length Polymomentioning

confidence: 99%

“…The tandem repetitions in T. gondii are often simple, with only repetitions of two nucleotides, which occur from 2 to 20 times [62,63]. Around 15 markers of MS have been used, including (tub2, W35, TgM-A, B18, B17; M33, IV.1, XI.1, M48, M102, N60, N82, AA, N61, N83) to determine the genotype of T. gondii in diferent laboratories [61,62]. Ajzenberg et al developed an easy method to genotype T. gondii in one multiplex PCR using 15 MS markers, in which eight MS markers (tub2, W35, TgM-A, B18, B17, M33, IV.1 and XI.1) could diferentiate the types I, II and III from all the atypical genotypes, and the other seven markers (M48, M102, N60, N82, AA, N61, N83 ) could improve the resolution from the genetic diferentiation of strictly related isolates in an haplo-group or clonal linage [62,63].…”

Section: Polymerase Chain Reaction-restriction Fragment Length Polymomentioning

confidence: 99%

“…Around 15 markers of MS have been used, including (tub2, W35, TgM-A, B18, B17; M33, IV.1, XI.1, M48, M102, N60, N82, AA, N61, N83) to determine the genotype of T. gondii in diferent laboratories [61,62]. Ajzenberg et al developed an easy method to genotype T. gondii in one multiplex PCR using 15 MS markers, in which eight MS markers (tub2, W35, TgM-A, B18, B17, M33, IV.1 and XI.1) could diferentiate the types I, II and III from all the atypical genotypes, and the other seven markers (M48, M102, N60, N82, AA, N61, N83 ) could improve the resolution from the genetic diferentiation of strictly related isolates in an haplo-group or clonal linage [62,63]. The multiplex 15-MS assay is the best available tool to characterize T. gondii from genetically diferent or identical isolates during an outbreak, laboratory contamination and from mixed infections.…”

Section: Polymerase Chain Reaction-restriction Fragment Length Polymomentioning

confidence: 99%

See 1 more Smart Citation

The Laboratory Diagnosis in Toxoplasma Infection

Ramírez¹,

Orozco²,

Ramírez³

2017

Toxoplasmosis

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The diagnosis of toxoplasmosis is of great importance due to the damage caused by this parasite in immunosuppressed people or in pregnant women, the diagnosis of an active toxoplasmosis represents a sign to initiate a pharmacological treatment immediately. The diagnosis of Toxoplasma can be performed with direct methods through intraperitoneal inoculation of serum or cerebrospinal luid, in susceptible mice evaluating the survival and detection of tachyzoites of biological samples. Indirect methods detecting the IgM and IgG isotypes against Toxoplasma have been the tools mostly used and had leaded to discriminate between an active and acute, from a chronic toxoplasmosis. Molecular methods actually Toxoplasma-DNA identiication by molecular biology tests like the polymerase chain reaction (PCR) allow the direct detection of the parasite. Polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLPs) have been used to identify three strain linages (type I, II and III). Recently, a high-resolution melting method was described to determine the genotype of the infection by Toxoplasma gondii directly from biological samples.

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Genetic variability of Brazilian Toxoplasma gondii strains detected by random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) and simple sequence repeat anchored-PCR (SSR-PCR)

Cited by 47 publications

References 60 publications

Efficacy of atovaquone and sulfadiazine in the treatment of mice infected withToxoplasma gondiistrains isolated in Brazil

Efficacy of atovaquone and sulfadiazine in the treatment of mice infected withToxoplasma gondiistrains isolated in Brazil

Characterization ofToxoplasma gondiifrom domestic animals from Minas Gerais, Brazil

The Laboratory Diagnosis in Toxoplasma Infection

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