荧光定量聚合酶链反应诊断弓形虫感染的实验研究  

Experimental study on the detection of toxoplasma DNA by fluorescence quantitative PCR techique

作　　者：符生苗[1] 阮和球[1] 陈鑫萍[1] 庞海云[1] 凌奕[1] 徐文[2] 杨力芳[2] 

机构地区：[1]海南省人民医院医学研究中心,海口市570311 [2]中山医科大学达安基因诊断中心,广州市510089

出　　处：《中国热带医学》2002年第1期6-8,共3页China Tropical Medicine

摘　　要：目的 探讨荧光定量聚合酶链反应（FQ-PCR）对孕妇与新生儿弓形虫（TOX）感染的诊断价值。方法以完全闭管式的PCR和荧光探针杂交技术相结合所产生的实时检测定量PCR方法,检测了产妇的血清标本和新生儿脐血标本,同时与常规 PCR法和酶联免疫吸附法（ELISA）比较。结果 225例产妇中,FQ-PCR阳性28例,平均拷贝数值（ml-1）为 1.68×105,阳性率为12.44％,常规PCR阳性率为12.89％,TOX-IgG阳性29例,阳性率为 12.89％,IgM阳性33例,阳性率为14.67％。179例新生儿中,FQ-PCR阳性7例,平均拷贝数值（ml-1）为2.29×105,阳性率为3.91％,常规PCR阳性率为5.03％,TOX-IgG阳性11例,阳性率为6.15％,IgM阳性3例,阳性率为1.68％。结论FQ-PCR特异性比ELISA法高,可以检测TOX的真实感染和复制情况,对TOX的临床诊断、治疗方案的选择和疗效观察有较大的应用价值。ve To study the diagnostic value of fluorescence quantitative PCR(FQ - PCR) technique for detection of toxoplasma in pregnant women and the new - borns and its clinical significance is also discussed . Methods Flrorescence - labeled probe hybridization in combination with PCR was used for quantitative detection toxoplasma DNA from the sera of the umbilical blood of pregnant women and the new - borns and comparisons with conventional PCR and ELISA were conducted. Results The Results of FQ - PCR shows that 28 cases have positive reaction out of 225 cases Lying - in woman serum samples(positive rate is 12.44%), the average DNA copies of positive samples was 1.68 × 105/ml; at the same time, 29 cases of TOX - IgG and 33 cases of TOX - IgM of these samples, positive rate is 12.89% and 14.61% . FQ - PCR shows that 7 cases have positive reaction out of 179 cases newborn infants umbilical blood samples(positive rate is 3. 91%) , the average DNA copies of positive samples was 2.29 × 105/ml; at the same time, 11 cases of TOX- IgG and 3 cases of TOX - IgM of these samples, positive rate is 6.15 % and 1.68% . Conclusion In - tube monitoring eliminates PCR cross -contamination which causes false positive, and real - time detection ensures accurate quantity. This method is also higher spectific. So FQ - PCR can be used to monitor the true state of TOX infection and amplification.

关 键 词：弓形虫 聚合酶链反应 DNA 弓形虫病 动物实验 诊断 TOX感染 

分 类 号：R531.8[医药卫生—内科学] R382.5[医药卫生—临床医学]