Development of a real-time recombinase-aided amplification (RT-RAA) molecular diagnosis assay for sensitive and rapid detection of Toxoplasma gondii
Development of a real-time recombinase-aided amplification (RT-RAA) molecular diagnosis assay for sensitive and rapid detection of Toxoplasma gondii
复制标题
开发实时重组酶辅助扩增 (RT-RAA) 分子诊断测定法，用于灵敏、快速检测弓形虫
DOI：
10.1016/j.vetpar.2021.109489
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发表时间：
2021
影响因子：
2.6
通讯作者：
Wang Xiao-Jia
中科院分区：
文献类型：
--
作者：
Wang Zhao-Hua;Zhang Wei;Zhang Xiu-Zhong;Yao Xin-Ran;Huang Wei;Jia Hong;Liu Xiao-Lei;Hou Shao-Hua;Wang Xiao-Jia
刚地弓形虫是一种细胞内寄生的原生动物，存在于广泛的宿主中，包括几乎所有种类的温血脊椎动物。弓形虫病通过各种途径传播给人类，包括受污染的食物或水，以及与各种家畜的密切接触。它对人类健康构成严重威胁，并造成重大经济损失，不仅在疾病成本方面，而且在监测方案方面。因此，有必要开发一种快速的现场即时诊断技术，以控制或防止病原体传播到经济上重要的牲畜、家畜和人类。在这项研究中，我们开发了一种实时等温扩增方法能够检测T。猪和猫血液样本中的弓形虫基因组。该方法在36 °C反应25 min，最低检测限为102 copies/reaction。与传统的实时荧光PCR相比，该方法具有灵敏度高、特异性强等优点，且可在便携式仪器上进行。
Toxoplasma gondii, a protozoan intracellular parasite, is present in a wide range of hosts, including virtually all species of warm-blooded vertebrates. Toxoplasmosis spreads to humans through a variety of pathways, including contaminated food or water, and close contact with various types of domestic animals. It poses a severe threat to human health, and contributes to important economic losses, not only in cost-of-illness but also in surveillance programs. It is thus necessary to develop a rapid point-of-care field diagnostic technology to control or prevent pathogen transmission to economically important livestock animals, domestic animals, and human beings. In this study, we develop a real-time isothermal amplification method capable of detecting theT. gondiigenome in swine and feline blood samples. This method can detecttoxoplasmagenome with a lowest detection limit of 102copies of per reaction under optimal reaction conditions of 36 °C for 25 min. The assay displayed advantages in sensitivity and specificity in comparison to traditional real-time PCR, and can be performed in a portable instrument.