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Experimental model of congenital toxoplasmosis in guinea-pigs: use of quantitative and qualitative PCR for the study of maternofetal transmission

Groupe Immunité des Muqueuses et Agents Pathogènes (GIMAP), Faculté de Médecine Jacques Lisfranc, 15 rue Ambroise Paré, 42023 Saint Etienne, France and *Unité d'Immunologie et de Physiologie, Département de Biologie, Faculté des Sciences et Techniques, Avenue A. El Khattabi, BP 618, 40000 Marrakech, Morocco

Corresponding author: Dr P. Flori (e-mail: pierre.flori{at}univ-st-etienne.fr).

Received 14 Dec. 2001; revised version received 3 April 2002; accepted 4 April 2002.

Maternofetal transmission of Toxoplasma gondii was assessed in pregnant guinea-pigs, with a gestational period of 65 ± 5 days. A total of 56 female guinea pigs was infected by the intraperitoneal route (RH strain), by the oral or the intraperitoneal route (Prugniaud strain; PRU) or by the oral route (76K strain). Inoculation was performed 90 ± 18 days or 30 ± 9 days before the onset of gestation or 20 ± 6 days or 40 ± 6 days after. Gestational age was determined by a progesterone assay. Parasite loads (fetal brain and liver) were assessed by nested PCR and real-time PCR quantification on Light Cycler^® was performed with a SYBR Green I^® technique. The 76K strain appeared to be the most virulent in the model: the neonatal survival rate was 31%, in contrast to 53% and 68% for the PRU and RH strains, respectively. The percentage of survival of neonates for all strains taken together was lower after inoculation at 40 days’ gestation (25%) than at 20 days’ gestation (77%). Whatever the strain, maternofetal transmission determination was greater with nested PCR (54% for RH, 84% for PRU and 86% for 76K strains) than with real-time quantitative PCR (31% for RH, 66% for PRU and 76% for 76K strains). However, real-time quantitative PCR showed that neonatal parasite load was greater with the cystogenic strains (76K, PRU) and that high hepatic load (>10 000 parasites/g) was often associated with disease severity (11 of 12 cases). Therefore, this technique may provide an important element in understanding this congenital disease.

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