Farid Alyaa, Aiad Silvia, Safwat Gehan
Biotechnology Department, Faculty of Science, Cairo University, Giza, Egypt.
Faculty of Biotechnology, October University for Modern Sciences and Arts (MSA), Giza, Egypt.
AMB Express. 2023 Nov 9;13(1):125. doi: 10.1186/s13568-023-01632-w.
Cryptosporidiosis is an intestinal infection that is triggered by the protozoan parasite Cryptosporidium spp. Cryptosporidium oocysts can spread from one host to another either through direct contact with infected hosts' faeces or through indirect means (consumption of contaminated water or food). Significant numbers of oocysts are produced as a result of the rapid growth of the parasite within the infected hosts. For proper care of cryptosporidiosis, a laboratory diagnosis is necessary. Therefore, this study aimed to produce anti-Cryptosporidium parvum (C. parvum) oocyst immunoglobulin (Ig)G polyclonal antibodies (pAbs). The produced pAbs were used in the detection of C. parvum oocysts antigens in stool and serum samples of infected calves. Moreover, pAbs were tested in protection of balb-c male mice from cryptosporidiosis infection. C. parvum oocysts were used in the preparation of antigens to be used in the immunization of New Zealand white rabbits. pAb was purified by ammonium sulphate precipitation method, caprylic acid purification method and diethylaminoethyl (DEAE) anion exchange chromatographic method. Sandwich enzyme-linked immunosorbent assay (ELISA) (using prepared pAb) scored higher sensitivity (85% and 95% for serum and stool samples) than that (80%) of microscopic examination of stool samples. Moreover, pAb significantly reduced the oocysts shedding, decreased inflammatory cytokines and enhanced the loss in the body weight of protected animals. The prepared pAb succeeded in the diagnosis of cryptosporidiosis in calves with high sensitivity either in the serum or stool samples. Our results indicated the usefulness of using the prepared pAb in protection against cryptosporidiosis.
隐孢子虫病是一种由原生动物寄生虫隐孢子虫属引发的肠道感染。隐孢子虫卵囊可通过直接接触受感染宿主的粪便或通过间接途径（食用受污染的水或食物）从一个宿主传播到另一个宿主。由于寄生虫在受感染宿主体内快速生长，会产生大量的卵囊。为了妥善治疗隐孢子虫病，实验室诊断是必要的。因此，本研究旨在制备抗微小隐孢子虫（C. parvum）卵囊免疫球蛋白（Ig）G多克隆抗体（pAbs）。所制备的多克隆抗体用于检测受感染犊牛粪便和血清样本中的微小隐孢子虫卵囊抗原。此外，还测试了多克隆抗体对雄性Balb-c小鼠免受隐孢子虫病感染的保护作用。微小隐孢子虫卵囊用于制备抗原，以免疫新西兰白兔。多克隆抗体通过硫酸铵沉淀法、辛酸纯化法和二乙氨基乙基（DEAE）阴离子交换色谱法进行纯化。夹心酶联免疫吸附测定（ELISA）（使用制备的多克隆抗体）的灵敏度（血清和粪便样本分别为85%和95%）高于粪便样本显微镜检查的灵敏度（80%）。此外，多克隆抗体显著减少了卵囊排出，降低了炎性细胞因子，并减轻了受保护动物的体重损失。所制备的多克隆抗体成功地以高灵敏度诊断了犊牛血清或粪便样本中的隐孢子虫病。我们的结果表明所制备的多克隆抗体在预防隐孢子虫病方面具有实用性。
