Zhang Hongbo, Zhang Nan, Li Jianhua, Zhao Panpan, Li Xin, Wang Xiaocen, Zhang Xu, Yuan Bao, Gao Fei, Gong Pengtao, Zhang Xichen
Key Laboratory of Zoonosis Research, Ministry of Education, College of Veterinary Medicine, Jilin University, Changchun 130062, China.
College of Animal Sciences, Jilin University, Changchun 130062, China.
Animals (Basel). 2023 Oct 11;13(20):3177. doi: 10.3390/ani13203177.
A variety of rodent ceca are parasitized by (), a flagellated protozoan. To date, there are no ideal methods for the detection of infections in laboratory mice; thus, new molecular methodologies for its specific detection need to be developed. In this study, using staining and SEM, it was observed that has a pear-shaped body and contains three anterior flagella. A nested PCR system with novel specific primers was designed based on the conserved regions of the SSU rRNA gene of . The nested PCR system for showed good specificity and high sensitivity for at least 100 trophozoites/mL and 0.1 ng/μL of fecal genomic DNA, which means that 176 trophozoites per gram of mouse feces could be detected. When using this nested PCR system, the detection rate was 18.96% (58/306), which was higher than the detection rate of 14.05% (43/306) detected via smear microscopy in fecal samples from five mouse strains. The sensitivity and specificity of nested PCR in detecting was found to be 100%, and it demonstrated a 26% increase in diagnostic sensitivity compared to the smear microscopy method in the present study. In conclusion, the nested PCR developed with novel primers based on the SSU rRNA gene of has good accuracy, specificity, and sensitivity for the detection of infections in laboratory mice.
多种啮齿动物的盲肠被一种鞭毛原生动物（）寄生。迄今为止，尚无理想的方法用于检测实验室小鼠中的感染；因此，需要开发用于其特异性检测的新分子方法。在本研究中，通过染色和扫描电子显微镜观察到，具有梨形身体并含有三根前鞭毛。基于的小亚基核糖体RNA（SSU rRNA）基因的保守区域设计了一种带有新型特异性引物的巢式PCR系统。针对的巢式PCR系统对每毫升至少100个滋养体和每微升0.1纳克粪便基因组DNA表现出良好的特异性和高灵敏度，这意味着每克小鼠粪便中176个滋养体可以被检测到。当使用该巢式PCR系统时，检测率为18.96%（58/306），高于通过涂片显微镜法在五个小鼠品系的粪便样本中检测到的14.05%（43/306）的检测率。发现巢式PCR检测的灵敏度和特异性均为100%，并且在本研究中与涂片显微镜法相比，其诊断灵敏度提高了26%。总之，基于的SSU rRNA基因开发的新型引物巢式PCR在检测实验室小鼠感染方面具有良好的准确性、特异性和灵敏度。
