重组小隐孢子虫CP2和CP23抗原在人隐孢子虫病血清诊断中的应用

Q2 Biochemistry, Genetics and Molecular Biology Iranian Biomedical Journal Pub Date : 2022-11-01 DOI:10.52547/ibj. 3801
Gholamreza Barzegar, Ehsan Ahmadpour, Bahador Shahriari, Rahmat Solgi, Mohammad Hossein Motazedian
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摘要

背景:小隐孢子虫是一种重要的球虫寄生虫,可感染包括人类在内的许多哺乳动物。这种寄生虫可在免疫功能低下的个体中表现为慢性严重腹泻,特别是艾滋病患者。本研究采用间接ELISA法,重组制备了小隐孢子虫的重组(r)P2和rP23抗原,作为检测人隐孢子虫病的抗原。方法:对rP2和rP23蛋白的编码序列进行密码子优化、商业合成并在pET28a表达载体上进行亚克隆。表达蛋白经Ni-NTA柱层析纯化,Western blotting证实。通过阳性(n = 20)和阴性(n = 20)人血清评估rP2/rP23蛋白的血清诊断效果,并通过Ziehl-Neelsen染色作为金标准试验来证实。结果:在ELISA检测中,小弓形虫感染患者血清对rP2/rP23反应强烈。ELISA检测中rP2/rP23与诊断潜力相关的敏感性和特异性均为100%。结论:ELISA中rP23和rP2抗原的结合可显著提高人隐孢子虫病的小隐孢子虫血清诊断效能。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

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Use of Recombinant CP2 and CP23 Antigens of Cryptosporidium parvum for Serodiagnosis of Human Cryptosporidiosis

Background: Cryptosporidium parvum is an important coccidian parasite infecting many mammals, including human. This parasite can manifest as chronic severe diarrhea in immunocompromised individuals, especially those with AIDS. The present study reports the recombinant production of recombinant (r)P2 and rP23 antigens of C. parvum as antigens for detecting human cryptosporidiosis using indirect ELISA tests.

Methods: The coding sequences of rP2 and rP23 proteins were codon-optimized, commercially synthesized and sub-cloned in the pET28a expression vector. The expressed proteins were purified by Ni-NTA column chromatography and confirmed by Western blotting. The efficacy of rP2/rP23 proteins for serodiagnosis was evaluated by positive (n = 20) and negative (n = 20) human sera, confirmed by the Ziehl-Neelsen staining as the gold standard test.

Results: In ELISA test, the sera from C. parvum-infected patients reacted strongly to rP2/rP23. The sensitivity and specificity related to the diagnostic potential of rP2/rP23 in the ELISA assay were 100%.

Conclusion: Our results showed that combination of rP23 and rP2 antigens in ELISA significantly increases the performance of C. parvum serodiagnosis in human cryptosporidiosis.

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来源期刊
Iranian Biomedical Journal
Iranian Biomedical Journal Biochemistry, Genetics and Molecular Biology-Biochemistry, Genetics and Molecular Biology (all)
CiteScore
3.20
自引率
0.00%
发文量
42
审稿时长
8 weeks
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