Real-time PCR targeting a 529-bp repeat element for diagnosis of toxoplasmosis.
Sensitive and rapid detection of infection with Toxoplasma gondii in transplanted immunocompromised patients is crucial for a good prognosis. Two DNA fragments are used currently for detecting T. gondii infection by PCR, i.e., the B1 gene and a 529-bp repeat element that exists in 200-300 copies/genome. This study investigated whether targeting the 529-bp repeat element gives better sensitivity and accuracy than can be obtained when targeting the B1 gene (35 copies) when concentrations of T. gondii DNA are low. The results demonstrated that detection of the 529-bp repeat element increased diagnostic sensitivity and accuracy. Addition of an internal amplification control did not affect the PCR performance and was useful in order to monitor PCR inhibition by non-specific DNA in the LightCycler instrument. The real-time PCR was used successfully in a clinical context to monitor parasitaemia in the blood of a transplant recipient suffering from toxoplasmosis.
Edvinsson
B
B
Karolinska Institutet, Department of Laboratory Medicine, Division of Clinical Bacteriology, Karolinska University Hospital Huddinge, Stockholm, Sweden. benjamin.edvinsson@labmed.ki.se
Lappalainen
M
M
Evengård
B
B
D000818:Animals; D001769:Blood / Q000469:parasitology; D016054:DNA, Protozoan / Q000032:analysis* / Q000235:genetics; D006801:Humans; D018512:Parasitemia / Q000175:diagnosis; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D011786:Quality Control; D012015:Reference Standards; D012091:Repetitive Sequences, Nucleic Acid / Q000235:genetics*; D012680:Sensitivity and Specificity; D014122:Toxoplasma / Q000235:genetics / Q000302:isolation & purification; D014123:Toxoplasmosis / Q000175:diagnosis*
D016054:DNA, Protozoan