Toxoplasma gondii DNA Detection

The buffy coat samples were used for DNA extraction using the phenol–chloroform–isoamyl alcohol method, as described in a previous study [16 (link)]. PCR was performed using the repetitive element (RE) gene amplifying a region of 529 base pairs (bp) fragments. The primers are highly sensitive and specific for T. gondii due to 200–300 replications in the T. gondii genome [17 (link)]. The PCR primers and cycling conditions were described in previous reports [18 (link), 19 (link)]. For each reaction, a positive control (DNA extracted from the RH strain of T. gondii) and a negative control (double-distilled water) were included. PCR products were electrophoresed in 2% agarose gel, stained with safe stain (Sinaclon, Iran), and visualized under a UV transilluminator.

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Haghbin M., Maani S., Bagherzadeh M.A., Bazmjoo A., Shakeri H., Taghipour A., Falahi S., Kenarkoohi A., Badri M, & Abdoli A. (2023). Latent Toxoplasmosis among Breast Cancer Patients in Jahrom, South of Iran. International Journal of Breast Cancer, 2023, 4792260.

Publication 2023

safe stain

Agarose Chloroform Gene Genome Isoamyl alcohol Phenol Primers Repetitive element Replications Stain Strain

Corresponding Organization : Jahrom University

Other organizations : Ilam University, Medical University of Ilam, Qazvin University of Medical Sciences

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Variable analysis

independent variables

DNA extraction method (phenol–chloroform–isoamyl alcohol)

dependent variables

Presence/absence of T. gondii DNA

control variables

PCR primers and cycling conditions

controls

Positive control (DNA extracted from the RH strain of T. gondii)
Negative control (double-distilled water)

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