Experimental model of congenital toxoplasmosis in guinea-pigs: use of quantitative and qualitative PCR for the study of maternofetal transmission
Experimental model of congenital toxoplasmosis in guinea-pigs: use of quantitative and qualitative PCR for the study of maternofetal transmission
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DOI：
10.1099/0022-1317-51-10-871
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发表时间：
2002-10-01
影响因子：
3
通讯作者：
Sung, RTM
中科院分区：
文献类型：
--
作者：
Flori, P;Hafid, J;Sung, RTM
在妊娠期为 65 +/- 5 天的怀孕豚鼠中评估了弓形虫的母胎传播情况。总共56只雌性豚鼠通过腹膜内途径（RH株）、口服或腹膜内途径（Prugniaud株；PRU）或口服途径（76K株）感染。接种在妊娠开始前90+/-18天或30+/-9天或后20+/-6天或40+/-6天进行。通过黄体酮测定确定孕龄。通过巢式 PCR 评估寄生虫负荷（胎儿大脑和肝脏），并使用 SYBR Green I(R) 技术在 Light Cycler(R) 上进行实时 PCR 定量。 76K 菌株似乎是模型中毒性最强的菌株：新生儿存活率为 31%，而 PRU 和 RH 菌株的新生儿存活率分别为 53% 和 68%。所有菌株的新生儿存活率在妊娠 40 天时接种后的存活率 (25%) 低于妊娠 20 天时的存活率 (77%)。无论何种菌株，巢式 PCR 的母胎传播测定结果（RH 为 54%，PRU 为 84%，76K 菌株为 86%）比实时定量 PCR（RH 为 31%，PRU 为 66%，76K 菌株为 76%）更高。然而，实时定量 PCR 显示，囊原菌株（76K，PRU）的新生儿寄生虫载量更大，并且高肝载量（>10 000 寄生虫/克）通常与疾病严重程度相关（12 例中有 11 例）。因此，这项技术可能为了解这种先天性疾病提供重要元素。
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(R) was performed with a SYBR Green I(R) 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.