Use of the polymerase chain reaction to detect Toxoplasma gondii in human blood samples.
OBJECTIVE
To assess the value of detecting Toxoplasma gondii in human blood samples using the polymerase chain reaction (PCR).


METHODS
Blood samples in lithium heparin were examined from 34 patients with suspected toxoplasmosis, and six healthy volunteers with or without the addition of doubling dilutions of toxoplasma tachyzoites. Products of a nested PCR, using oligonucleotide primers of the B1 gene, were analysed by electrophoresis and stained by ethidium bromide. The primary product was 194 base pairs in length; the nested products were 160 or 97 base pairs.


RESULTS
When toxoplasma tachyzoites were added to the leucocytes of six different volunteers, eight to 16 parasites were detected by nested PCR in one experiment and one to four parasites in eight experiments. All nine experiments were negative in samples without added tachyzoites. Of 34 patients, PCR was negative in 13 with recent lymphadenopathy; nine with persisting IgM, including two pregnant patients; four with reactivated infection due to immunodeficiency; and five with no evidence of active infection. Positive PCR results were found in three patients with reactivated infection. There was only one discrepancy between PCR and animal culture results; this was in an immunocompromised patient with a positive PCR and negative culture.


CONCLUSIONS
The PCR technique was rapid, sensitive, and specific in human blood samples. Negative PCR results in patients with persisting IgM suggested that the fetus was not at risk, or that treatment was not indicated in myalgic encephalomyelitis-like illness. PCR results in immunocompromised patients permitted appropriate management--no treatment if negative, treatment if positive.
Ho-Yen
D O
DO
Scottish Toxoplasma Reference Laboratory, Raigmore Hospital, Inverness.
Joss
A W
AW
Balfour
A H
AH
Smyth
E T
ET
Baird
D
D
Chatterton
J M
JM
D000818:Animals; D005260:Female; D006801:Humans; D007075:Immunoglobulin M / Q000032:analysis; D007962:Leukocytes / Q000382:microbiology; D016133:Polymerase Chain Reaction / Q000379:methods; D011247:Pregnancy; D014122:Toxoplasma / Q000302:isolation & purification; D014123:Toxoplasmosis / Q000175:diagnosis*
D007075:Immunoglobulin M