State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan 430072, China.
Acta Trop. 2011 Oct-Nov;120(1-2):88-94. doi: 10.1016/j.actatropica.2011.06.006. Epub 2011 Jul 8.
Schistosomiasis is a major infectious disease and a public health concern in many areas in China and other countries. Sensitive method for detection of the parasite is critical for early diagnosis and for monitoring of effective treatment of the disease. In this study, we developed a highly sensitive TaqMan real-time PCR assay for the detection of Schistosoma japonicum DNA in mouse feces and serum samples. This assay was based on the DNA sequence of the S. japonicum 18S rRNA gene and was able to detect 10 fg of S. japonicum genomic DNA, which is 100 times more sensitive than conventional PCR. We were able to detect the S. japonicum DNA one week post-infection in mouse sera and 4 weeks post-infection in feces, which was one week earlier than egg detection by microscopy in feces. This assay was also highly specific for Asian Schistosomes which are causative species of human Schistosomiasis. In single sex male cercariae infected mice, parasite DNA was only detected in the first 4 weeks post-infection, suggesting that the DNA was derived from decaying worms' corpse in the first 4 weeks whereas the DNA was mainly from decaying parasite eggs afterwards. Therefore we conclude that the established TaqMan real-time PCR assay is a sensitive, specific and convenient method that could be used for the early diagnostic evaluation of S. japonicum infection in humans and for monitoring outbreaks in endemic areas with low prevalence.
血吸虫病是中国和其他国家许多地区的主要传染病和公共卫生关注点。寄生虫的敏感检测方法对于早期诊断和监测疾病的有效治疗至关重要。在这项研究中,我们开发了一种高度敏感的 TaqMan 实时 PCR 检测方法,用于检测小鼠粪便和血清样本中的日本血吸虫 DNA。该检测方法基于日本血吸虫 18S rRNA 基因的 DNA 序列,能够检测到 10 fg 的日本血吸虫基因组 DNA,比常规 PCR 灵敏 100 倍。我们能够在感染后一周的小鼠血清中检测到日本血吸虫 DNA,在感染后 4 周的粪便中检测到,比粪便中显微镜检测到虫卵早一周。该检测方法也高度特异性地针对亚洲血吸虫,亚洲血吸虫是人类血吸虫病的致病物种。在感染单性雄性尾蚴的小鼠中,仅在感染后 4 周内检测到寄生虫 DNA,表明在第 1-4 周内,DNA 来自死亡虫体的残骸,而之后 DNA 主要来自死亡的寄生虫卵。因此,我们得出结论,建立的 TaqMan 实时 PCR 检测方法是一种敏感、特异和方便的方法,可用于人类日本血吸虫感染的早期诊断评估,并监测低流行地区的暴发。