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一种先进的尿嘧啶DNA糖基化酶补充型环介导等温扩增(UDG-LAMP)技术,用于艾滋病患者中微小隐孢子虫、人隐孢子虫和火鸡隐孢子虫的灵敏特异检测。

An advanced uracil DNA glycosylase-supplemented loop-mediated isothermal amplification (UDG-LAMP) technique used in the sensitive and specific detection of Cryptosporidium parvum, Cryptosporidium hominis, and Cryptosporidium meleagridis in AIDS patients.

作者信息

Fallahi Shirzad, Moosavi Seyedeh Fatemeh, Karimi Azadeh, Chegeni Ali Sharafi, Saki Mohammad, Namdari Parsa, Rashno Mohammad Menati, Varzi Ali Mohamad, Tarrahi Mohammad Javad, Almasian Mohammad

机构信息

Razi Herbal Medicine Research Center, Lorestan University of Medical Sciences, Khorramabad, Iran; Department of Medical Parasitology and Mycology, Faculty of Medicine, Lorestan University of Medical Sciences, Khorramabad, Iran.

Department of Medical Parasitology and Mycology, Faculty of Medicine, Lorestan University of Medical Sciences, Khorramabad, Iran.

出版信息

Diagn Microbiol Infect Dis. 2018 May;91(1):6-12. doi: 10.1016/j.diagmicrobio.2017.12.017. Epub 2017 Dec 24.

Abstract

The rapid and accurate detection of Cryptosporidium spp. is critically important for the prevention and timely treatment of cryptosporidiosis in AIDS patients (APs). This study was conducted to examine a UDG-LAMP technique for the first time to diagnose cryptosporidiosis in APs. After collecting demographic and clinical data, three stool samples were collected from the participants (120 volunteering APs). The microscopic examination of stained smears using the acid-fast method and the UDG-LAMP assay were performed for each sample. 10% of APs were infected with Cryptosporidium spp. The number of detected cryptosporidiosis cases using the acid-fast staining and UDG-LAMP methods were significantly different (P < 0.001). Diarrhea and weight loss were found to be significantly associated with cryptosporidiosis in patients (P < 0.05). The pretreatment of LAMP reagents with UDG successfully eliminated the likelihood of product re-amplification remaining from previous reactions. The UDG-LAMP technique could detect cryptosporidiosis in APs with high sensitivity and rapidity without carryover contamination.

摘要

快速准确地检测隐孢子虫属对于艾滋病患者(APs)隐孢子虫病的预防和及时治疗至关重要。本研究首次采用UDG-LAMP技术诊断艾滋病患者的隐孢子虫病。收集人口统计学和临床数据后,从参与者(120名自愿参与的艾滋病患者)中采集了三份粪便样本。对每个样本进行抗酸染色涂片的显微镜检查和UDG-LAMP检测。10%的艾滋病患者感染了隐孢子虫属。使用抗酸染色和UDG-LAMP方法检测到的隐孢子虫病病例数有显著差异(P < 0.001)。腹泻和体重减轻被发现与患者的隐孢子虫病显著相关(P < 0.05)。用UDG对LAMP试剂进行预处理成功消除了先前反应残留产物再次扩增的可能性。UDG-LAMP技术能够高灵敏度、快速地检测艾滋病患者的隐孢子虫病,且无交叉污染。

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