Mugasa Claire M, Schoone Gerard J, Ekangu Rosine A, Lubega George W, Kager Piet A, Schallig Henk D F H
Faculty of Veterinary Medicine, Department of Veterinary Parasitology and Microbiology, Makerere University, Kampala, Uganda.
Diagn Microbiol Infect Dis. 2008 Aug;61(4):440-5. doi: 10.1016/j.diagmicrobio.2008.03.019. Epub 2008 May 16.
Currently, the conventional diagnosis of human African trypanosomiasis (HAT) is by microscopic demonstration of trypomastigotes in blood, lymph, and/or cerebrospinal fluid. However, microscopic diagnosis of HAT is not sensitive enough and may give false-negative results, thus, denying the patient the necessary treatment of the otherwise fatal disease. For this reason, a highly sensitive technique needs to be developed to enhance case findings. In this study, the real-time nucleic acid sequence-based amplification assay described is based on amplification and concurrent detection of small subunit rRNA (18S rRNA) of Trypanosoma brucei. The sensitivity of the assay was evaluated on nucleic acid from in vitro cultured parasites and blood spiked with various parasites quantities. The assay detected 10 parasites/mL using cultured parasites as well as spiked blood. A sensitive assay such as the one developed in this study may become an alternative tool to confirm diagnosis of human African trypanosomiasis.
目前，人类非洲锥虫病（HAT）的传统诊断方法是通过显微镜检查血液、淋巴液和/或脑脊液中的锥鞭毛体。然而，HAT的显微镜诊断不够灵敏，可能会得出假阴性结果，从而使患者无法得到对这种可能致命疾病的必要治疗。因此，需要开发一种高灵敏度技术以加强病例发现。在本研究中，所描述的基于实时核酸序列的扩增检测方法是基于布氏锥虫小亚基核糖体RNA（18S rRNA）的扩增及同步检测。该检测方法的灵敏度通过体外培养寄生虫的核酸以及加入不同数量寄生虫的血液进行评估。使用培养的寄生虫以及加样血液时，该检测方法能检测出每毫升10个寄生虫。像本研究中开发的这种灵敏检测方法可能会成为确诊人类非洲锥虫病的替代工具。
