Repetitive DNA sequences of Toxoplasma gondii for development of diagnostic probes

Angel, Sérgio O.; Blanco, Jorge C. G.; Maero, E; Pzsenny, Viviana; Garberi, Juan C.

doi:10.1590/s0074-02761991000400023

Mem. Inst. Oswaldo Cruz

1991

DOI: 10.1590/s0074-02761991000400023

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Repetitive DNA sequences of Toxoplasma gondii for development of diagnostic probes

Sérgio O. Angel

Jorge C. G. Blanco

E Maero

et al.

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Cited by 4 publications

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“…Aliquots of CSF samples were spotted onto a nitrocellulose filter and processed as described elsewhere (5). Hybridization conditions and DNA immunodetection were as described previously (1). The presence of mycobacterium and fungi was investigated in aliquots of the same CSF samples by specific culture.…”

mentioning

confidence: 99%

Early diagnosis of toxoplasmic encephalitis in AIDS patients by dot blot hybridization analysis

et al. 1992

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Central nervous system toxoplasmosis is a life-threatening infection with a mortality rate of higher than 60%. An early and rapid diagnosis is important for effective treatment of the disease. A new approach for detection of cerebral toxoplasmosis is described here. DNAs extracted from cells in cerebrospinal fluid samples (0.3 to 0.8 ml) of patients suspected of having cerebral toxoplasmosis were analyzed by a dot blot hybridization technique. A highly repetitive DNA sequence of Toxoplasma gondii (ABGTg4) was nonisotopically labelled with digoxigenin-dUTP and used as a specific DNA probe. Four of six patients analyzed gave positive signals in our hybridization assay. Two of them recovered with pyrimethamine-sulfadiazine, a drug recommended for treatment of toxoplasmosis. The other two patients with positive signals died soon after diagnosis. Patients with negative signals were found to suffer from mycobacterial infection (patient 1) or varicella-zoster virus infection (patient 6).

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mentioning

confidence: 99%

Early diagnosis of toxoplasmic encephalitis in AIDS patients by dot blot hybridization analysis

et al. 1992

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Echinococcus granulosus:Cloning and Characterization of a Tandemly Repeated DNA Element

Rosenzvit

Canova

Kamenetzky

et al. 1997

Experimental Parasitology

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Detection of Toxoplasma gondii parasitemia by gene amplification, cell culture, and mouse inoculation

Hitt

Filice

1992

J Clin Microbiol

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Diagnosis of Toxoplasma gondii infection is difficult, especially in immunosuppressed people. The AIDS epidemic has increased the number of people at risk and increased the need for better diagnostic methods. We have compared three methods for detection of T. gondii parasitemia. Rabbits were infected subcutaneously with 104 T. gondii tachyzoites. Blood samples were obtained, and buffy coat or leukocyte fractions were prepared. We sought the T. gondii Bi gene by gene amplification by the polymerase chain reaction, and we sought viable T. gondii cells by inoculating fibroblast cell cultures and by mouse inoculation. Thirty-two blood samples were obtained from seven infected rabbits, and 18 were obtained from four control, uninfected rabbits. Parasitemia was detected in 20 of 32 samples (62%) from infected samples by mouse inoculation, 12 of 32 samples (37%) by gene amplification and detection, and 8 of 32 samples (25%) by cell culture. Mouse inoculation requires use of live animals and has a long turnaround time. Currently, cell culture is the least sensitive but most practical and widely available method for the detection of T. gondii parasitemia. Gene amplification and detection was more sensitive than cell culture and may become available in clinical laboratories as techniques are developed further and automated.

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Repetitive DNA sequences of Toxoplasma gondii for development of diagnostic probes

Cited by 4 publications

References 0 publications

Early diagnosis of toxoplasmic encephalitis in AIDS patients by dot blot hybridization analysis

Early diagnosis of toxoplasmic encephalitis in AIDS patients by dot blot hybridization analysis

Echinococcus granulosus:Cloning and Characterization of a Tandemly Repeated DNA Element

Detection of Toxoplasma gondii parasitemia by gene amplification, cell culture, and mouse inoculation

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