绿脓杆菌感染肺炎大鼠模型中的PCR检测方法及IL23的动态变化  

Application of PCR Detection in the Rats with Pseudomonas Aeruginosa Pneumonia and the Dynamic Changes of IL23 in Rats

作　　者：乌兰.同巴依尔 颜婕 刘媛媛[1] 牛晓琳[1] 王志强[1] 白煜峡[1] Wulan·tongbayier;YAN Jie;LIU Yuan-yuan;NIU Xiao-lin;WANG Zhi-qiang;BAI Yu-xia(First Affiliated Hospital of Xinjiang Medical University:Disinfecting supply division,Ummqi,Xinjiang,830054,China;First Affiliated Hospital of Xinjiang Medical University:Central operatingroom,Ummqi,Xinjiang,830054,China)

机构地区：[1]新疆医科大学第一附属医院消毒配送中心次供应室一部,新疆乌鲁木齐830054 [2]新疆医科大学第一附属医院中心手术室,新疆乌鲁木齐830054

出　　处：《现代生物医学进展》2018年第23期4416-4420,共5页Progress in Modern Biomedicine

基　　金：新疆维吾尔自治区重点实验室医学动物模型研究实验室开放课题(XJDX1103-2012-08)

摘　　要：目的:探讨PCR技术在绿脓杆菌感染肺炎大鼠模型中进行检测的方法可行性以及大鼠体内IL23的表达水平变化。方法:将健康SD大鼠随机分为3组,空白对照组、实验对照组和肺炎感染组。在绿脓杆菌肺炎模型建立后,设计绿脓杆菌的特异性引物,进行3组大鼠体内绿脓杆菌感染的PCR鉴定,并用qPCR和ELISA技术检测大鼠体内IL23水平变化。结果:①PCR技术检测到肺炎大鼠模型中的绿脓杆菌,其中肺组织检出率为100%,而对照组的PCR结果是阴性。②绿脓杆菌感染肺炎大鼠模型建立后,大鼠的IL23 m RNA逐渐上升,在第3天达到最高值,相比对照组,有显著差异(分别是60852.9±1474.2和1987.7±246.4拷贝,P<0.05);血清中的IL23蛋白水平也逐渐上升,在第1周到达最高值,相比对照组,有显著差异(分别是297.4±14.7和123.8±18.3pg/m L,P<0.05)。结论:本研究建立了大鼠体内的高特异性、快速、简便绿脓杆菌感染检测PCR方法,发现大鼠IL23在绿脓杆菌感染后显著升高,在绿脓杆菌肺炎和IL23之间建立了联系。Objective: To explore the feasibility of PCR detection and the change of IL23 expression level in the rat model of pyocyanic pneumonia. Methods: Healthy SD rats were randomly divided into three groups, blank control group, experimental control group and pneumonia group. After the establishment of pyocyanic pneumonia model, specific primers of Pseudomonas aeruginosa were designed, and PCR was carried out to identify whether the rats were infected or not. The IL23 level in rats was detected by qPCR and ELISA. Results:①Pseudomonas aeruginosa was detected in the rat with Pseudomonas aeruginosa pneumonia by PCR, and the positive ratio was 100% in lung, but the results in the control group were negative.②After the establishment of pyocyanic pneumonia model, the content of IL23 m RNA showed gradual increases, and reached the top level on the third day. When compared to the control group, there was significant difference(60852.9 ±1474.2 and 1987.7 ±246.4 copies, respectively, P<0.05). The content of serum IL23 increased gradually, and reached the top level in the first week. When compared to the control group, there was significant difference(297.4±14.7and 123.8±18.3 pg/mL, respectively, P<0.05). Conclusions: A highly specific, rapid and simple PCR method for detection of Pseudomonas aeruginosa infection in rats was established. IL23 in rats increased significantly after infection with Pseudomonas aeruginosa, and a connection between pyocyanic pneumonia and IL23 was established.

关 键 词：绿脓杆菌 肺炎 PCR检测 IL23 

分 类 号：R-33[医药卫生] R378