结核病生物诊断试剂豚鼠模型建立

Establishment of a guinea pig model for biological diagnostic reagent of tuberculosis

目的建立结核病生物诊断试剂模型。方法通过在豚鼠腹股沟皮下单次和多次注射不同剂量灭活H37Rv全菌体,制备结核病生物诊断试剂模型,采用0.1 mL(5 IU)标准结核菌素(TB-PPD)对模型效果进行评价。结果三组剂量(0.2 mg/mL,0.3 mg/mL和0.5 mg/mL)致敏豚鼠均能建立模型,其中0.2 mg/mL剂量组皮试后24 h和48 h红斑纵横径值均大于0.3 mg/mL和0.5 mg/mL剂量组,致敏效果最好;多次致敏皮试红斑纵横径值与单次致敏比较无显著差异;第1次皮试后24 h红斑纵横径值大于第2次、第3次和第4次皮试后红斑纵横径值(P≤0.05);此外,灭活H37Rv全菌体致敏后,豚鼠体重持续增加,部分豚鼠注射部位有溃烂发生。结论应用0.2mg/mL灭活H37Rv全菌体单次致敏豚鼠,可以建立良好的用于结核病生物诊断试剂评价的模型。增加致敏剂量和致敏次数不能增加变态反应强度。

Objective To establish a guinea pig model for diagnostic reagent of tuberculosis.Methods By single or multiple subcutaneous injection of heat-killed H37 Rv in different doses in the groin of guinea pigs to establish a model of positive response to 0.1 mL （5 IU） standard tuberculin （ TB-PPD） skin test.Results Three doses of heat-killed H37 Rv （ 0.2 mg/mL, 0.3 mg/mL and 0.5 mg/mL ） could be used to generate the model of biological diagnosis of tuberculosis.After 24 and 48 hours, the diameter of red spot by TB-PPD skin test was 15.4 ±2.3 mm when a dose of 0.2 mg/mL heat-killed H37 Rv was administered for immunizing and allergizing the guinea pigs.The biggest red spot was induced at doses of 0.3 mg/mL and 0.5 mg/mL.The test results showed that the immune response induced by multiple njection to immunizing and allergizing guinea pigs was not significantly different than that induced by single immunizing injection, and the first skin test was better than the second, third and fourth skin test （P≤0.05）.In addition, the body weight of the guinea pigs was still increasing after infection with heat-killed H37 Rv, and ulcers occurred in the injection sites in some guinea pigs.Conclusions A single subcutaneous injection of 0.2 mg/mL heat-killed H37 Rv in guinea pigs can be used well to establish a reliable model for biological diagnostic reagent of tuberculosis.Increasing the sensitizing dose and multiple sensitization can not increase the intensity of the delayed-type hypersensitivity （ DTH） response.