关键词: 弓形虫; PCR检测试剂盒; 组装; 应用

Abstract: To develop PCR detection kit for Toxoplasma and observe the effects of clinical applicaytion in this study. The MIC3 gene sequences of Toxoplasma were chosen to design the species-specific primers on GenBank. The PCR amplification conditions were optimized and a series of tests were conducted respectively in the specificity, sensitivity, stability, reproducibility, and stored period of the kit, and expanded clinical trials were also carried out. This kit could specifically amplify MIC3 gene the expected DNA fragment of Toxoplasma. There was no cross-reaction with other parasites, such as Coccidian, Giardia, Trichinella and Cryptosporidium. It was able to detect as low as 10 of Toxoplasma gondii tachyzoites DNA. The results of different batches of kits of the same sample were consistent, and had higher repeatability and stability. The kit worked well after being stored at room temperature for at least twelve months. Its effect of clinical application was significant. The results turned out that this PCR detection kit could provide a means of early diagnosis, epidemiological investigation and quarantine of Toxoplasma.

Key words: Toxoplasma; PCR detection kit; development; application