Effect of rSAG-1(P30) immunisation on the circulating and tissue parasites in guinea pigs as determined by quantitative PCR.

The efficacy of immunisation with Toxoplasma gondii recombinant protein (rSAG-1) was evaluated in the guinea pig model. For the infectious challenge, two strains, namely, strain C56 (10,000 tachyzoites) and strain 76K (100 cysts), were used to infect a group of 32 guinea pigs each. The circulating, cerebral and pulmonary parasite loads were determined with the real-time polymerase chain reaction (PCR) after immunisation. With the C56 strain, immunisation showed high activity with a reduction of greater than 1 log of the circulating and tissue parasite loads. Thus, there was a significantly lower circulating parasite load in the rSAG-1 + adjuvant group (0.5+/-1.5 Eq parasites/ml) as compared to that in the control group (67+/-110 Eq parasites/ml; p<0.05). In the same manner, the cerebral parasite load was much lower in the rSAG-1 + adjuvant group (10+/-20 Eq parasites/g) than that in the control group (339+/-291 Eq parasites/g; p<0.01). On the other hand, with the 76K strain, the effect of immunisation was much less and that only on the pulmonary parasite load [p(lung)<0.05]. This could be due to the use of different strains and stages of the parasite and/or the difference in the route of administration for challenge. The quantitative PCR technique used has shown a good correlation with animal inoculation, and when associated with the guinea pig model, it seems to be a useful and reproducible technique for future vaccine studies.

Reliability of immunoglobulin G antitoxoplasma avidity test and effects of treatment on avidity indexes of infants and pregnant women.

The immunoglobulin G antitoxoplasma avidity test (Vidas; BioMérieux) is an immunoenzymatic test useful for excluding acute infection after the onset of pregnancy. The avidity index (AI) is the ratio of the signal in a test sample washed with urea, which disrupts low-avidity complexes, to that washed without urea. An AI of >0.3 is taken to mean that infection had occurred more than 4 months ago. The increase of the AI with time and the influence of the different treatments given to pregnant women and their newborns were evaluated. A total of 59 pregnant women (271 sera) and their 60 neonates (199 sera) were tested from 1998 to 2002. There were five groups of women based on the type and duration of treatment given. Thirteen pregnant women (group 1) did not receive any treatment, 15 (group 2), 11 (group 3), and 17 (group 4) women received treatment with spiramycin (9 MIU/day) for 0.5 to 2, 2.5 to 5, and 5.5 to 8 months, respectively, and the last 3 women (group 5) received tritherapy (pyrimethamine-sulfonamide and spiramycin alternatively) for 1.5 to 2.5 months. All of the maternal sera collected in the first 6 months had an AI of <0.30, with a mean of 0.07 (range, 0.01 to 0.21). The increase was slow (0.02/month), and there was no significant difference when comparisons were made between the treatment groups. Neonates with proven maternofetal transmission had an increasing AI, unlike those without transmission. However, long-term therapy with pyrimethamine-sulfonamide, as opposed to treatment with spiramycin alone, was found to slow down the progression of the AI. An AI of >0.2 is sufficient to exclude acute infection in pregnant women. In neonates, it is not of major use to diagnose congenital infection; however, it could be a good indicator of compliance and efficacy of treatment of infected infants.