摘要
目的应用 PCR 检验技术在口岸进行鼠疫检测,以提高鼠疫诊断的准确性和灵敏度,建立适合于口岸鼠疫检测的快速诊断方法,提高鼠疫检测水平。方法首先,根据口岸地区样品的特点确定相应 PCR 检测方法,然后对样品进行 PCR 检测、鼠疫细菌学检验和反向间接血球凝集试验。结果形成了一套适合于口岸地区鼠疫 PCR 检测的方法,灵敏度为7cfu 的鼠疫菌。PCR 反应体系中以 dUTP 代替 DTTp,加入 UDG 酶防止产物污染,对特异性和敏感性无影响。对在口岸地区采集的鼠类标本111份、进口旱獭皮张500份进行了 PCR 检测,并分别进行鼠疫细菌学、反向间接血球凝集试验。PCR 检测出3份阳性鼠类标本,与鼠疫细菌学检验结果一致:对进口旱獭皮张的 PCR 检测和反向间接血球凝集试验均为阴性。结论经实验研究建立了一套完整的鼠疫 PCR 检测方法。本方法特异性和敏感性强、操作简便、快捷、成本较低,适合在口岸地区推广应用。
Objective Appliance PCR technique to the ditection of plague at port region,and increases the veracity and delicacy of plague detection.To establishing a method of speediness diagnosis that suited for port plague detection improve the level of plague's detection at port region.Methods First,According to the characters of samples at port region,we make sure an appropriate method of PCR detection,and then proceed to these samples with PCR detection,Plague bacteriology examine and reversed indirection hemagglutination(RIHA). Results We accomplished a suit of method of plague PCR detection.Delicacy of the method is 7cfu plague germs.In PCR reaction system,dTTP is replaced with dUTP,and mixed with UDG emzyme to defend pollution of PCR production have no influence to the sensitivity and the particularity.We have examined 111 murine specimens by PCR and Plague bacteriology methods.PCR method examined 3 positive specimens, the result is consistent with Plague bacteriology methods.We have examined 500 pieces Marmot skin by PCR and RIHA methods,the specimen are negative in all.Conclusion By experimented and studied,we have established a suit of complete PCR detection method which suit for plague detection at port region.
