Int Conf AIDS. 1993 Jun 6-11;9(1):386 (abstract no. PO-B10-1503). Unique
OBJECTIVE: To study Toxoplasma gondii (Tg) DNA amplification in the
cerebrospinal fluid (CSF) of AIDS patients (pts). MATERIALS AND METHODS:
Frozen aliquots of CSF and serum from 3 AIDS pts. with brain lesions
recovered after anti-Toxo therapy. Samples were obtained just before
start of therapy in 2 pts. and one week after in the third (sample C).
DNA was phenol-chloroform extracted from 150 microliters of CSF;
alternatively, 20 microliters of CSF were boiled for 10 min then
directly used as a template for PCR; 55 cycles of PCR were done with
primers flanking the BI 35-folds repetitive sequence of Tg genome. PCR
products were hybridizated with a biotinilated probe, in a microplate
Elisa system (DEIA, Sorin, Italy). A nested PCR was also performed, with
inner primers located inside the first pair; PCR products were subjected
to gel electrophoresis and ethidium bromide staining. Negative controls
were used at each phase of the procedure. RESULTS: the 3 CSF were
positive after both nested PCR and DEIA; sample C was weakly positive
after DEIA and was negative by both methods after rapid DNA extraction.
Sera were PCR negative in the 3 cases. CONCLUSIONS: PCR for Tg DNA in
the CSF may aid diagnosis of toxo-encephalitis in AIDS pts. Correlation
to clinical and neuroradiological pictures must be established.
*DNA, Protozoan/CEREBROSPINAL FLUID *Encephalitis/ETIOLOGY
*Toxoplasma/ISOLATION & PURIF *Toxoplasmosis, Cerebral/DIAGNOSIS