Duplex quantitative real-time PCR assay for the detection and discrimination of the eggs of Toxocara canis and Toxocara cati (Nematoda, Ascaridoidea) in soil and fecal samples.
Duplex quantitative real-time PCR assay for the detection and discrimination of the eggs of Toxocara canis and Toxocara cati (Nematoda, Ascaridoidea) in soil and fecal samples.
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DOI：
10.1186/1756-3305-5-288
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发表时间：
2012-12-07
影响因子：
3.2
通讯作者：
Gala JL
中科院分区：
文献类型：
--
作者：
Durant JF;Irenge LM;Fogt-Wyrwas R;Dumont C;Doucet JP;Mignon B;Losson B;Gala JL
弓蛔虫病是由犬弓蛔虫（T. canis）和/或猫弓蛔虫（T. cati）引起的人畜共患疾病，这两种世界各地分布的蛔虫，分别是犬科动物和猫科动物的寄生虫。人类感染是通过在玩耍被狗或猫粪便污染的土壤时摄入犬毛虫或猫毛虫的胚胎卵而引起的。因此，评估这些蛔虫卵对这些区域的潜在污染至关重要。针对核糖体 RNA 基因内转录间隔区 (ITS2) 的双重定量实时 PCR (2qPCR) 已开发出来，可用于快速、特异性地鉴定粪便和土壤样品中的犬毛蛔虫和猫毛蛔虫卵。该检测采用从 53 条成虫中提取的 DNA 样本，包括犬蛔虫、猫蛔虫、狮子蛔虫、猪蛔虫 (A. suum) 和马蛔虫 (P. equorum)。该检测用于评估多个样品中是否存在猫蛔虫卵，包括 12 个掺有猫蛔虫卵或猪蛔虫卵的干净土壤样品、从布鲁塞尔游乐场随机收集的 10 个实际土壤样品，以及来自猫、狗和其他动物的粪便样本。将狗和猫粪便样本的 2qPCR 结果与显微镜检查结果进行比较。 2qPCR 检测可以特异性检测犬毛虫和猫毛虫，无论是成虫、土壤中的虫卵还是粪便样本。加标土壤样品中的 2qPCR 检测限 (LOD) 为每克土壤 2 个鸡蛋，周转时间为 3 小时。对狗和猫的粪便进行 2qPCR 检测和显微镜检查之间观察到完美的一致性。新开发的 2qPCR 检测可用于高通量前瞻性或回顾性检测粪便样本以及游乐场、公园和沙坑土壤样本中的犬锥虫和/或猫锥虫卵。
Toxocarosis is a zoonotic disease caused by Toxocara canis (T. canis) and/or Toxocara cati (T. cati), two worldwide distributed roundworms which are parasites of canids and felids, respectively. Infections of humans occur through ingestion of embryonated eggs of T. canis or T. cati, when playing with soils contaminated with dogs or cats feces. Accordingly, the assessment of potential contamination of these areas with these roundworms eggs is paramount. A duplex quantitative real-time PCR (2qPCR) targeting the ribosomal RNA gene internal transcribed spacer (ITS2) has been developed and used for rapid and specific identification of T. canis and T. cati eggs in fecal and soil samples. The assay was set up on DNA samples extracted from 53 adult worms including T. canis, T. cati, T. leonina, Ascaris suum (A. suum) and Parascaris equorum (P. equorum). The assay was used to assess the presence of T. cati eggs in several samples, including 12 clean soil samples spiked with eggs of either T. cati or A. suum, 10 actual soil samples randomly collected from playgrounds in Brussels, and fecal samples from cats, dogs, and other animals. 2qPCR results on dogs and cats fecal samples were compared with results from microscopic examination. 2qPCR assay allowed specific detection of T. canis and T. cati, whether adult worms, eggs spiked in soil or fecal samples. The 2qPCR limit of detection (LOD) in spiked soil samples was 2 eggs per g of soil for a turnaround time of 3 hours. A perfect concordance was observed between 2qPCR assay and microscopic examination on dogs and cats feces. The newly developed 2qPCR assay can be useful for high throughput prospective or retrospective detection of T.canis and/or T. cati eggs in fecal samples as well as in soil samples from playgrounds, parks and sandpits.
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影响因子：
2.6
作者：
Deplazes, Peter;van Knapen, Frans;Overgaauw, Paul A. M.
通讯作者：
Overgaauw, Paul A. M.
影响因子：
2.7
作者：
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通讯作者：
Chilton, NB
影响因子：
1.6
作者：
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通讯作者：
Mizgajska-Wiktor, H.
影响因子：
11.8
作者：
Bouchaud, O;Houzé, S;Coulaud, JP
通讯作者：
Coulaud, JP
影响因子：
3.2
作者：
Chen J;Xu MJ;Zhou DH;Song HQ;Wang CR;Zhu XQ
通讯作者：
Zhu XQ