Polymerase chain reaction for detection of Toxoplasma gondii

Savva, D.; Morris, Judy C.; Johnson, Julie; Holliman, R E

doi:10.1099/00222615-32-1-25

Cited by 120 publications

(55 citation statements)

References 16 publications

(7 reference statements)

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“…Encephala samples, as well as lungs and hearts from GIC, displayed a considerable positivity, and the highest rate was observed for the thigh muscles. These results agree with the data shown in studies done with fresh tissues of mice chronically infected by T. gondii, which show a consistent positiveness in brain samples (Savva et al 1990, Owen & Trees 1998, Jauregui et al 2001, as well as in heart and skeletal musculature (Homan et al 2000). DNA of the parasite was detected by PCR and hybridization assays in the same kind of tissues that were positive for the immunohistochemistry analysis, including lungs, revealing the ability of the technique in detecting the infection by T. gondii in CIG.…”

Section: Resultssupporting

confidence: 90%

Detection of Toxoplasma gondii DNA by polymerase chain reaction in experimentally desiccated tissues

Terra

Bello

Bastos

et al. 2004

Mem. Inst. Oswaldo Cruz

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Section: Resultssupporting

confidence: 90%

Detection of Toxoplasma gondii DNA by polymerase chain reaction in experimentally desiccated tissues

Terra

Bello

Bastos

et al. 2004

Mem. Inst. Oswaldo Cruz

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“…Detection of T. gondii was carried out using nested PCR amplification of the Surface Antigen Gene 1 (Savva et al 1990) as modified by Morley et al (2005). The PCR product from one positive example of each species was extracted from the gel using the GENECLEAN 2 kit (Q-BIOgene) and sequenced in both directions (Lark Technologies Inc.) to confirm the detection of Toxoplasma (Accession nos.…”

Section: Pcr Detection Of Toxoplasma Gondiimentioning

confidence: 99%

The prevalence ofNeospora caninumand co-infection withToxoplasma gondiiby PCR analysis in naturally occurring mammal populations

et al. 2005

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Neospora caninumandToxoplasma gondiiare closely related intracellular protozoan parasites associated with bovine and ovine abortion respectively. Little is known about the extent ofNeospora/Toxoplasmaco-infection in naturally infected populations of animals. Using nested PCR techniques, based on primers from the Nc5 region ofN. caninumand SAG1 forT. gondii, the prevalence ofN. caninumand its co-infection withT. gondiiwere investigated in populations ofMus domesticus,Rattus norvegicusand aborted lambs (Ovis aries). A low frequency of infection withN. caninumwas detected in theMus domesticus(3%) andRattus norvegicus(4·4%) populations. A relatively high frequency of infection withN. caninumwas detected in the brains of aborted lambs (18·9%). There was no significant relationship betweenN. caninumandT. gondiico-infection. Investigation of the tissue distribution ofNeospora, in aborted lambs, showed thatNeosporacould not be detected in tissues other than brain and this was in contrast toToxoplasmawhere the parasite could be frequently detected in a range of tissues.

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“…SAG1 gene (GenBank -X14080) was amplified according to previously described methods [16] and SAG4 gene (GenBank -Z69373) amplification was based on primer pairs derived from Ö dberg-Ferragut et al with modified conditions of PCR to increase the stringency [22].…”

Section: Laboratory Conditions For Pcrmentioning

confidence: 99%

“…This may account for the failure of PCR which targets the B1 gene [6][7][8][9][10][11]. Recently, cloning of stage-specifically expressed genes, such as SAG1, located in the tachyzoites, or BAG1, SAG4 and MAG1, located in the bradyzoites, has enabled further analysis of regulatory mechanisms that are involved in the stage interconversion and provided insights into the pathogenesis of toxoplasmosis [3,[12][13][14][15][16][17][18].…”

Section: Introductionmentioning

confidence: 99%

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

Contini¹,

Cultrera

Seraceni³

et al. 2002

Journal of Medical Microbiology

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The switch from bradyzoites to tachyzoites is the fundamental pathogenic event that leads to Toxoplasma gondii encephalitis (TE) in patients with AIDS. Distinction between these stages is difficult, particularly when specific treatment has been started. A new approach consisting of a nested PCR (n-PCR) assay was performed on cerebrospinal fluid (CSF) specimens collected from AIDS patients with TE before or after antiparasitic therapy was initiated, to assess the efficacy of primer sets which amplify target sequences expressed on bradyzoites (SAG4 and MAG1), tachyzoites (SAG1) or both stages (B1) of T. gondii. CSF specimens were obtained from 46 patients with AIDS, of whom 27 had TE (16 first episode, 11 relapse) and 19 had other AIDS-related brain lesions (AIDS-OBL) in the absence of TE. CSF specimens from 26 HIV-negative and immunocompetent patients were also checked. All samples were tested with different primer pairs targeting the B1, SAG-1, SAG-4 and MAG-1 genes. With B1, 75% of patients with first episodes of TE were positive, compared with 36.3% of those with relapse of TE and 5.2% of those with AIDS-OLB. The SAG1 gene yielded positive values in 28.7% and 45.4% of patients with first episodes of TE or relapse of TE, respectively, and in none of the controls. With the SAG4 and MAG1 genes, 72.7% of patients with relapse of TE were detected, compared with 25% of patients with first episodes of TE and 5.2% with AIDS-OLB. None of the HIV-negative subjects showed positive PCR reactions. These results demonstrate that specific primers for the genes SAG4, MAG1 and SAG1 may be useful in AIDS patients with relapse of TE, in whom the use of PCR targeting the B1 gene may fail to detect DNA, especially when prophylaxis or treatment has been started.

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Polymerase chain reaction for detection of Toxoplasma gondii

Cited by 120 publications

References 16 publications

Detection of Toxoplasma gondii DNA by polymerase chain reaction in experimentally desiccated tissues

Detection of Toxoplasma gondii DNA by polymerase chain reaction in experimentally desiccated tissues

The prevalence ofNeospora caninumand co-infection withToxoplasma gondiiby PCR analysis in naturally occurring mammal populations

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

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