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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 To detect the plague in the port by using the PCR test technique to improve the accuracy and sensitivity of the plague diagnosis and to establish a rapid diagnostic method suitable for the plague detection at the port so as to improve the plague detection level. Methods Firstly, the corresponding PCR detection method was determined according to the characteristics of the samples in the port area, then the PCR test, the plague bacteriological test and the reverse indirect hemagglutination test were carried out on the samples. Results A set of methods for PCR detection of plague in the port area was developed, with a sensitivity of 7 cfu Yersinia pestis. PCR reaction system instead of DTTp dUTP, UDG enzyme added to prevent product contamination, specificity and sensitivity. 111 samples of rodents collected in the port area and 500 copies of imported marmot were detected by PCR and plague bacteriology and indirect hemagglutination test respectively. Three positive samples were detected by PCR, which was consistent with the results of the plague bacteriological test. PCR tests and reverse indirect hemagglutination test were all negative for imported marmot hide. Conclusion The experimental study established a complete set of plague PCR detection methods. The method is specific and sensitive, easy to operate, fast and low in cost, and is suitable for popularization and application in the port area.