卡氏肺孢子菌肺炎大鼠中的LAMP检测方法及1,3-β-D葡聚糖变化的研究

摘要

目的 探讨环介导恒温扩增法(LAMP)检测在卡式肺孢子菌肺炎大鼠中的可行性及1,3-β-D葡聚糖水平的变化.方法 40只Wister大鼠随机分为对照组和肺炎感染组,每组各20只.采用地塞米松磷酸钠构建卡式肺孢子菌肺炎大鼠模型.模型构建成功后,观察两组大鼠的发病情况、体质量、肺重及肺重体积比.设计卡式肺孢子菌的特异度引物,对两组大鼠进行LAMP检测.采用酶联免疫吸附试验(ELISA)法检测外周血和肺灌洗液1,3-β-D葡聚糖水平.结果 与对照组比较,肺炎感染组大鼠死亡4只,体质量明显降低,肺重及肺重体积比明显上升,差异有统计学意义(P＜0.05);LAMP方法能检测卡式肺孢子菌,对照组为阴性;与对照组比较,肺炎感染组外周血和肺灌洗液1,3-β-D葡聚糖水平升高,且肺灌洗液1,3-β-D葡聚糖水平高于外周血,差异有统计学意义(P＜0.05).结论 本研究成功构建卡式肺孢子菌肺炎大鼠模型并建立卡式肺孢子菌简便快捷的LAMP检测方法,1,3-β-D葡聚糖水平与卡式肺孢子菌有一定的相关性.%Objective To investigate the feasibility of LAMP detection in Pneumocystis carinii pneumonia rats and the changes of 1, 3-β-D-glucan.Methods 40 Wister rats were randomly divided into control group and lung infection group, 20 rats in each group.Specific primers were designed for pneumoniae carinii and LAMP technique was carried out to identify whether the rats were infected or not.The levels of 1, 3-β-D-glucan in peripheral blood and lung lavage fluid were detected by ELISA.Results Compared with the control group, there were 4 dead rats in the lung infection group, the body weight decreased significantly, and the lung weight and the percentage of the lung volume increased significantly (P＜0.05).LAMP method can detect Pneumocystis carinii, the control group was negative.Compared with the control group, the level of 1, 3-β-D-glucan in peripheral blood and lung lavage fluid in the lung infection group increased.And the 1, 3-β-D-glucan level in lung lavage fluid was higher than that in peripheral blood (P＜0.05).Conclusion In this study, we successfully constructed a rat model of Pneumocystis carinii pneumonia and established a simple and rapid method for LAMP detection of Pneumocystis carinii.1, 3-β-D-glucan and Pneumocystis carinii have some relevance.