Rong Yao, Zhang Xichen, Chen Xuejiao, Li Jianhua, Gong Pengtao, Wang Xiaocen, Li Xin, Zhang Xu, Yue Taotao, Zhang Hongbo, Zhou Xiaofei, Zhang Nan
Key Laboratory of Zoonosis Research of Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine, Jilin University, Changchun 130062, China.
Curr Issues Mol Biol. 2023 Nov 17;45(11):9252-9261. doi: 10.3390/cimb45110579.
is a trichomonad protozoan that infects the cecum and colon of humans and other mammals. It is a zoonotic pathogen that causes diarrhea in both animals and humans. As companion animals, dogs infected with pose a risk of transmitting it to humans. Current methods, such as direct smears and polymerase chain reaction (PCR), used for detection have limitations, including low detection rates and the need for specialized equipment. Therefore, there is an urgent need to develop rapid, sensitive, and simple detection methods for clinical application. Recombinase polymerase amplification (RPA) has emerged as a technology for rapid pathogen detection. In this study, we developed a lateral flow dipstick (LFD)-RPA method based on the highly conserved gene for detecting infection by optimizing the primers, probes, and reaction conditions, and evaluating cross-reactivity with genomes of and other parasites. The LFD-RPA method was then used to test 128 dog fecal samples collected from Changchun. The results confirmed the high specificity of the method with no cross-reactivity with the five other parasites. The lowest detection limit of the method was 10 copies/µL, and its sensitivity was 10 times higher than that of the conventional PCR method. Consistent with the positivity rate observed using nested PCR, 12 samples (out of 128) tested positive using this method (positivity rate, 9.38%). In conclusion, the LFD-RPA method developed in this study represents a simple and sensitive assay that allows for the rapid detection of infection in dogs, especially in this field.
是一种毛滴虫原生动物,可感染人类和其他哺乳动物的盲肠和结肠。它是一种人畜共患病原体,可导致动物和人类腹泻。作为伴侣动物,感染 的狗有将其传播给人类的风险。目前用于 检测的方法,如直接涂片和聚合酶链反应(PCR),存在局限性,包括检测率低和需要专门设备。因此,迫切需要开发用于临床应用的快速、灵敏且简单的检测方法。重组酶聚合酶扩增(RPA)已成为一种快速病原体检测技术。在本研究中,我们通过优化引物、探针和反应条件,并评估与 和其他寄生虫基因组的交叉反应性,开发了一种基于高度保守的 基因的侧向流动试纸条(LFD)-RPA方法,用于检测 感染。然后使用LFD-RPA方法对从长春采集的128份犬粪便样本进行检测。结果证实了该方法具有高特异性,与其他五种寄生虫无交叉反应。该方法的最低检测限为10拷贝/µL,其灵敏度比传统PCR方法高10倍。与使用巢式PCR观察到的阳性率一致,使用该方法检测的128份样本中有12份(阳性率为9.38%)呈阳性。总之,本研究开发的LFD-RPA方法是一种简单且灵敏的检测方法,可快速检测犬中的 感染,尤其是在该领域。