Helmy Moshira M F
Department of Parasitology, Medical College, Misr University for Science and Technology, 6th October City, Egypt.
J Egypt Soc Parasitol. 2007 Dec;37(3):903-13.
A detection assay for Schistosoma mansoni DNA in mouse serum samples based on touchdown PCR was evaluated. Fifty four mice (7 weeks old) were infected with 60 +/- 10 cercariae of S. mansoni (Egyptian strain) through tail skin. Three days and 1-8 weeks post-infection (PI), six mice were randomly chosen from the infected ones and blood was collected by cardiac puncture. Sera and faeces were collected for 8 weeks PI. Antisoluble egg antigen (SEA) IgG for S. mansoni was detected in infected mice sera of at 3 weeks PI by ELISA. The earliest deposition of eggs in the small intestines was at the 5th week PI. The eggs were detected in faeces at the 8th weeks by modified Kato's thick-smear. However, S. mansoni DNA was detected by touchdown PCR in sera at 2 weeks PI. The intensity of S. mansoni DNA bands in infected mice sera detected in polyacrelamide gel electrophoresis increased gradually during the experimental period. Data recommended touchdown PCR potential tool for the early diagnosis of S. mansoni infection.
评估了一种基于降落式聚合酶链反应（touchdown PCR）的小鼠血清样本中曼氏血吸虫DNA检测方法。54只7周龄小鼠通过尾皮感染60±10条曼氏血吸虫（埃及株）尾蚴。感染后3天及感染后1至8周，从感染小鼠中随机选取6只，通过心脏穿刺采血。在感染后8周收集血清和粪便。通过酶联免疫吸附测定（ELISA）在感染小鼠感染后3周的血清中检测到针对曼氏血吸虫的抗可溶性虫卵抗原（SEA）IgG。最早在感染后第5周在小肠中发现虫卵沉积。在感染后第8周通过改良加藤厚涂片法在粪便中检测到虫卵。然而，通过降落式聚合酶链反应在感染后2周的血清中检测到曼氏血吸虫DNA。在聚丙烯酰胺凝胶电泳中检测到的感染小鼠血清中曼氏血吸虫DNA条带强度在实验期间逐渐增加。数据表明降落式聚合酶链反应是曼氏血吸虫感染早期诊断的潜在工具。
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