Application of the polymerase chain reaction to the diagnosis of human toxoplasmosis.
Toxoplasmosis may cause significant damage to the developing fetus and is a life-threatening opportunistic infection in immunocompromised persons. Serological investigation is unreliable, while isolation of the parasite is time consuming and may lack sensitivity. We have developed a system for detecting Toxoplasma gondii based on the amplification of the P30 gene using sequential rounds of PCR and nested primers. The clinical value of this technique was assessed by the investigation of a range of tissues taken from pregnant women, fetuses, neonates, AIDS patients and organ graft recipients. The PCR assay produced more positive reactions than isolation of the parasite by means of cell culture or animal inoculation. Extended autoradiography was found to be more sensitive than stained agarose gels for detecting the PCR product. Systematic contamination of PCR reactions was avoided but it was not possible to exclude sporadic contamination in certain cases. Detection of specific DNA is of clinical value in the investigation of the pregnant woman in order to assess the risk of transplacental passage of infection and in the fetus and neonate to identify congenital toxoplasmosis. Even so, PCR findings must be interpreted with caution because of the risk of a sample being contaminated. PCR may be the investigation of choice when brain biopsy is performed on a patient with AIDS and when toxoplasmosis associated with bone marrow transplantation is suspected.
Johnson
J D
JD
Toxoplasma Reference Laboratory, St. George's Hospital Medical School, London, U.K.
Butcher
P D
PD
Savva
D
D
Holliman
R E
RE
D017088:AIDS-Related Opportunistic Infections / Q000175:diagnosis; D000818:Animals; D016026:Bone Marrow Transplantation / Q000276:immunology; D016054:DNA, Protozoan / Q000032:analysis; D005260:Female; D005315:Fetal Diseases / Q000175:diagnosis; D006801:Humans; D007121:Immunocompetence; D007231:Infant, Newborn; D009894:Opportunistic Infections / Q000469:parasitology; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D015597:Pregnancy Complications, Parasitic / Q000175:diagnosis; D014122:Toxoplasma / Q000302:isolation & purification; D014123:Toxoplasmosis / Q000175:diagnosis* / Q000469:parasitology; D016781:Toxoplasmosis, Cerebral / Q000175:diagnosis; D014125:Toxoplasmosis, Congenital / Q000175:diagnosis
D016054:DNA, Protozoan