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NLM AIDSLINE

Polymerase chain reaction (PCR) for Toxoplasma gondii DNA detection in the cerebrospinal fluid of AIDS patients with acute Toxoplasma encephalitis.




 

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



 




Information in this article was accurate in November 30, 1993. The state of the art may have changed since the publication date. This material is designed to support, not replace, the relationship that exists between you and your doctor. Always discuss treatment options with a doctor who specializes in treating HIV.