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經(jīng)小鼠尾靜脈注射誘導(dǎo)肺炎克雷伯菌血流感染模型的建立及評價

發(fā)布時間:2018-03-21 02:32

  本文選題:肺炎克雷伯菌 切入點:毒力因子 出處:《中華醫(yī)院感染學(xué)雜志》2017年06期  論文類型:期刊論文


【摘要】:目的建立可靠的肺炎克雷伯菌血流感染模型,為細菌性血流感染早期診斷相關(guān)研究提供可靠的依據(jù)。方法選擇肺炎克雷伯菌標(biāo)準(zhǔn)株ATCC 700603為實驗菌株,將不同濃度的細菌懸液通過尾靜脈注入小鼠體內(nèi),按Karber法計算LD50,以1/2LD50濃度注射小鼠,通過其體征變化、體重變化、血培養(yǎng)、分子生物學(xué)試驗、H·E染色等指標(biāo)綜合評價模型,驗證造模效果。結(jié)果小鼠LD50為1.11×109 CFU/ml,菌液感染小鼠1h后開始出現(xiàn)立毛,活動減少;感染前體重為(33.60±1.21)g,注射菌液后1天內(nèi)體重下降至(28.86±1.42)g,跟對照組比較有明顯差別(P0.05),7天后恢復(fù)正常水平(33.17±1.72)g;正常小鼠白細胞為(1.42±0.66)×109/L,注射菌液3h后升至(2.52±1.01)×109/L,6h升至(3.08±0.85)×109/L,跟對照組比較有明顯差別(P0.05),2天后白細胞進一步升至(6.88±3.11)×109/L,7天后仍無明顯下降;小鼠全血培養(yǎng)結(jié)果表明前2天任意時間血培養(yǎng)結(jié)果為陽性;毒力因子引物擴增疑似菌DNA,結(jié)果表明小鼠血流感染病原菌均為肺炎克雷伯菌;注射菌液1天后小鼠肺泡明顯滲出、水腫、白細胞浸潤;2天后肺結(jié)構(gòu)逐漸恢復(fù),肝恢復(fù)速度較肺慢。結(jié)論建立了小鼠肺炎克雷伯菌血流感染模型,能夠為血流感染的早期診斷和不同病原菌感染的鑒別診斷等研究提供了可靠的動物模型。
[Abstract]:Objective to establish a reliable blood stream infection model of Klebsiella pneumoniae, and to provide reliable basis for the early diagnosis of bacterial blood flow infection. Methods ATCC 700603, a standard strain of Klebsiella pneumoniae, was selected as the experimental strain. Different concentrations of bacterial suspensions were injected into mice via tail vein, LD50 was calculated by Karber method, and 1 / 2 LD50 was injected into mice. A comprehensive evaluation model was established based on the changes of physical sign, body weight, blood culture, molecular biology test and H 路E staining. Results the LD50 of mice was 1.11 脳 109CFU / ml. The body weight before infection was 33.60 鹵1.21g, and the body weight dropped to 28.86 鹵1.42g within 1 day after injection of bacterial solution, which was significantly different from that of the control group, and returned to normal level after 7 days (33.17 鹵1.72g). The leukocyte of normal mice was 1.42 鹵0.66 脳 109 / L, and rose to 2.52 鹵1.01 脳 109L / L and 3.08 鹵0.85 脳 109L / L after 3 h injection of bacterial solution, and 3.08 鹵0.85 脳 109L / L after 3 h injection of bacterial solution, compared with the control group, the white blood cells of normal mice were 1.42 鹵0.66 脳 109 / L, respectively. After 2 days, the leukocyte level increased to 6.88 鹵3.11) 脳 10 ~ 9 / L ~ (-1) and no significant decrease was found after 7 days. The results of whole blood culture of mice showed that the results of blood culture were positive at any time of the first 2 days; the DNA of suspected bacteria was amplified by the primers of virulence factor. The results showed that the pathogenic bacteria of blood stream infection in mice were Klebsiella pneumoniae. After 2 days of edema, leukocyte infiltration, the lung structure gradually recovered, and the recovery rate of liver was slower than that of lung. Conclusion the blood stream infection model of Klebsiella pneumoniae was established in mice. It can provide a reliable animal model for the early diagnosis of blood flow infection and the differential diagnosis of different pathogens.
【作者單位】: 溫州醫(yī)科大學(xué)檢驗醫(yī)學(xué)院生命科學(xué)學(xué)院;解放軍總醫(yī)院臨床檢驗科;解放軍總醫(yī)院微生物科;
【基金】:國家重大科學(xué)儀器專項項目任務(wù)(2012YQ18011708)
【分類號】:R-332;R515

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