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使用免疫层析法分离少量微小隐孢子虫卵囊。

Isolation of Small Number of Cryptosporidium parvum Oocyst Using Immunochromatography.

作者信息

Ebrahimzade Elahe, Shayan Parviz, Asghari Zeinab, Jafari Sedighe, Omidian Zahra

机构信息

Department of Parasitology, Iranian Center for Ticks and Tick-borne Diseases, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran.

出版信息

Iran J Parasitol. 2014 Oct-Dec;9(4):482-90.

Abstract

BACKGROUND

Cryptosporidium parvum causes severe gastroenteritis in immunocompromised human and new borne animals. The organism can be transmitted through water. Since small number of C. parvum is infectious, the aim of the present study was to develop a chromatography method for the isolation of C. parvum oocyst in samples with limited number of oocysts.

METHODS

Antibody was prepared against whole antigen from C. parvum oocysts, the achieved Ab bound to the sepharose 4B and used for the isolation of oocysts. Antibody against P23 bound to the sepharose 4B, used also for the isolation of C. parvum oocyst. In comparison to these both methods, 2 traditional methods (Salt floatation and 55% sucrose floatation) were also performed.

RESULTS

Both chromatography methods could bind oocysts with capacity depends on the column size. The isolated oocysts were free of bacteria. Our results showed that the traditional methods are useful for the isolation of oocysts from feces, in its smear stained with ziehl-nelsen, at least 3 oocyts are detectable in each microscopic field under 1000 X magnification. In contrast to the chromatography methods, the bacterial contamination was always observed in oocysts isolated with traditional methods.

CONCLUSION

Immunochromatography could be used for the successful isolation of C. parvum oocysts from the samples containing limited number of oocysts.

摘要

背景

微小隐孢子虫可导致免疫功能低下的人和新生动物发生严重的肠胃炎。该病原体可通过水传播。由于少量的微小隐孢子虫即具有传染性,因此本研究的目的是开发一种色谱方法,用于从卵囊数量有限的样本中分离微小隐孢子虫卵囊。

方法

制备针对微小隐孢子虫卵囊全抗原的抗体,将获得的抗体与琼脂糖4B结合,用于卵囊的分离。针对P23的抗体与琼脂糖4B结合,也用于微小隐孢子虫卵囊的分离。与这两种方法相比,还采用了2种传统方法(盐浮选法和55%蔗糖浮选法)。

结果

两种色谱方法均可结合卵囊,结合能力取决于柱的大小。分离出的卵囊不含细菌。我们的结果表明,传统方法对于从粪便中分离卵囊很有用,在其经齐-尼氏染色的涂片中,在1000倍放大倍数下每个显微镜视野中至少可检测到3个卵囊。与色谱方法不同,用传统方法分离的卵囊中总是观察到细菌污染。

结论

免疫色谱法可成功地从卵囊数量有限的样本中分离微小隐孢子虫卵囊。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9f8e/4345087/205067df021a/IJPA-9-482f1.jpg

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