弓形体感染小鼠试验及基因快速诊断

IDENTIFICATION OF TOXOPLASMA GONDII IN CLINICAL SAMPLES AND EXPERIMENTAL MICE BY POLYMERASE CHAIN REACTION

为建立检测弓形体基因(Tox-DNA)的PCR快速诊断法.采用10只感染弓形体小鼠试验剖检取得的各脏器活检组织标本,32例畸形胎儿的脑、肺、肝、胎盘标本,29例正常产妇胎盘标本,及177例孕产妇和其它传染病人血液标本,用PCR法作了Tox-DNA测定.结果10只感染弓形体小鼠的脑、心、肺、肝、脾、肾、小肠、子宫/睾丸,腹水中均检出Tox-DNA.32例畸形胎儿的脑、肺、肝、胎盘中检出Tox-DNA 10例,阳性率31.25%,29例正常产妇胎盘对照Tox-DNA阳性1例,阳性率3.4%,两者比较差异有显著性,P<0.005.177例孕产妇及其它传染病人Tox-DNA阳性7例,阳性率3.95%.结论:小鼠感染弓形体可通过血液和淋巴侵犯各个脏器组织,引起病理性损害;孕产妇感染弓形体是致胎儿畸形的一个重要因素;PCR检测临床标本中Tox-DNA具有简便、快速、敏感、特异之优点,可用于弓形体病的早期诊断.

The aim of this study was to detect toxoplasma gene in various biological specimens for the diagnosis of T.gondii infection. Specific T. gondii gene amplification by PCR using the primers were 5'-GGAACTGCATCCGT-TCATGAG- 3' and 5'-TCTTTAAAGCGTTCGTGGTC-3'. The primers are located on the BI gene. We were able todetect T. gondii by PCR in clinical specimens and tissue samples of experimental mice. Autopsy samples of various tissuesobtained from 10 experimental mice infected with T. gondii, 128 tissues samples obtained from 32 fetal anomaly,and 177clinical specimens obtained from pregnant women and other infection patients, and 29 placenta tissues samples obtainedfrom normaly parturient control were detected for presence of toxoplasma gene(Tox-DNA) by PCR. The results of Tox-DNA detected in various biological specimens showed that the presence of T. gondii was demonstrated in brain, heart,lung,liver,spleen,kidney,uterus/testis.enteron,and scitic fluid obtained from 10 experimental mice infected with T. gondii, 10(31.25%) of 32 fetal anomaly,7(3. 95%) of 177 pregnant women and other infection patients, 1(3. 4%) of 29normaly parturient control. We are suggested the PCR can be used as a diagnostic tool for the rapid detection of T. gondiiin various clinical materials.