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应用环介导等温扩增(lamp)技术对流浪猫进行弓形虫种属的分子评估,作为一种快速、敏感、简单的筛选检测方法。

Molecular evaluation of Toxocara species in stray cats using loop-mediated isothermal amplification (lamp) technique as a rapid, sensitive and simple screening assay.

机构信息

Department of Pathobiology, Faculty of Veterinary Medicine, Lorestan University, Khorramabad, Iran.

Hepatitis Research Center, Lorestan University of Medical Science, Khorramabad, Iran.

出版信息

Vet Med Sci. 2021 May;7(3):647-653. doi: 10.1002/vms3.431. Epub 2021 Jan 25.

Abstract

Toxocara species are parasitic nematodes of dogs and cats with a worldwide distribution. The adult worm lives in the intestine, and horizontal transmission of the infection occurs through eating paratenic host or embryonated eggs. This study aimed to estimate the molecular prevalence of Toxocara species in stray cats using the loop-mediated isothermal amplification (LAMP) technique. A total of 95 stool samples were randomly collected from stray cats in Khorramabad city in western Iran. Microscopic examination was performed after the separation and extraction of supernatants. The LAMP reaction was performed using the internal transcribed spacer 2 (ITS2) gene primers of Toxocara species and the appropriate master mix. The overall prevalence of Toxocara spp. in stray cats was 20% (19/95, CI 95%: 0.2 ± 0.08) by parasitological and molecular assessments. The microscopic examination of stool samples revealed that 19 samples were positive for Toxocara. The same 19 positive samples were also positive by the LAMP technique. Interestingly, based on the results of the LAMP assay, out of 95 studied samples, 18 (18.94%; CI 95%: 0.19 ± 0.08) specimens were Toxocara canis, while only 1 (1.05%; CI 95%: 0.005 ± 0.01) sample was diagnosed as Toxocara cati. The relatively high prevalence of Toxocara species in the studied cats shows the role of this species in spreading the parasite and the role of the cats in transmitting this zoonotic parasite. Preventive measures including the control of stray cat's population by castration and protection of public gardens where children play are recommended. The easy, highly sensitive and specific LAMP method is proposed for the differential detection of Toxocara species in animals and humans.

摘要

弓形虫种类是犬和猫的寄生线虫,分布于世界各地。成虫生活在肠道中,感染的水平传播通过食用中间宿主或胚胎化卵发生。本研究旨在使用环介导等温扩增(LAMP)技术估计流浪猫中弓形虫种类的分子流行率。从伊朗西部霍拉马巴德市的流浪猫中随机采集了 95 份粪便样本。在分离和提取上清液后进行显微镜检查。使用 Toxocara 种的内部转录间隔区 2(ITS2)基因引物和适当的主混合物进行 LAMP 反应。通过寄生虫学和分子评估,流浪猫中弓形虫属的总流行率为 20%(19/95,95%置信区间:0.2±0.08)。粪便样本的显微镜检查显示,19 个样本对弓形虫呈阳性。同样,19 个阳性样本也通过 LAMP 技术呈阳性。有趣的是,根据 LAMP 检测结果,在所研究的 95 个样本中,18 个(18.94%;95%置信区间:0.19±0.08)标本为犬弓首蛔虫,而仅有 1 个(1.05%;95%置信区间:0.005±0.01)样本被诊断为猫弓首蛔虫。在所研究的猫中,弓形虫种类的相对高流行率表明该物种在传播寄生虫方面的作用,以及猫在传播这种人畜共患寄生虫方面的作用。建议采取包括绝育控制流浪猫数量和保护儿童玩耍的公共花园等预防措施。建议使用简单、高度敏感和特异的 LAMP 方法来检测动物和人类中的弓形虫种类。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/442a/8136963/12740f27d9b3/VMS3-7-647-g006.jpg

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