摘要
目的建立敏感、特异、稳定的聚合酶链反应（ＰＣＲ）法，以检测弓形虫感染。方法建立ＰＣＲ法，确定最佳扩增条件；用ＰＣＲ法检测实验感染家兔血液中ＤＮＡ的动态变化，并与ＥＬＩＳＡ法检测ＣＡｇ进行比较。结果兔感染弓形虫后第２ｄ，ＰＣＲ开始出现阳性，阳性率为７６．９％（１０／１３），总检出率达１００％。与ＥＬＩＳＡ法查ＣＡｇ进行平行检测，前者出现阳性时间早，且阳性率明显高于后者。该法敏感性高，可检测到１０ｆｇＤＮＡ含量；特异性强，对其它９种寄生虫和微生物ＤＮＡ均无交叉现象；稳定性好，对阳、阴性同一样品重复５次，结果一致。结论提示本法可用于临床诊断，在一定范围内替代病原学和免疫学检测。
Aim:To establish a sensitive,specific and stable PCR technique for detecting toxoplasma infection in experimental rabbits Method\ PCR technique was established and different amplified conditions were compared to detect the dynamic variation of serum DNA of toxoplasma in experimental infected rabbits then using the technigue at the same time to detect CAg with ELISA method Results\ DNA of toxoplasma could be detected as early as the second day from 10 out of the 13 rabbits after infection with a positive rate of 76 9% At the 30th day,the positive rate reached the highest value 91 7%(11/12) The total detection rate of toxoplasma DNA was 100% In comparison with CAg detecting by ELISA parallelly performed,this method is more sensitive for as little as 10fg of DNA could be detected Besides,the positive results could be detected earlier and the positive rate was higher than that of ELISA This method is also very specific No corss reaction was found with the DNA of nine other parasites and microorganisms Identical results were obtained when a positive or a negative sample was tested repeatedly for five times,showing that this method is quite stable and reliable Conclusion\ PCR can be used for clinical diagnosis of toxoplasmosis and to some extent,it can replace the conventional immunological and pathogenical examinations
出处
《中国人兽共患病杂志》
CSCD
北大核心
1999年第2期27-31,共5页
Chinese Journal of Zoonoses
关键词
弓形体
聚合酶链反应
家兔
Toxoplasam\ Polymerase chain reaction\ Experimentally infected rabbit
