發(fā)布時間：2018-07-09 15:31

  本文選題：鮑曼不動桿菌 + 生物被膜��； 參考：《中國人民解放軍醫(yī)學院》2013年碩士論文

【摘要】：1、目的：（1）回顧性分析我院2010-2012年鮑曼不動桿菌（AB）所致老年患者醫(yī)院獲得性肺炎（HAP）病例，總結臨床特點、易患因素及耐藥表型，為臨床合理應用抗菌藥物提供參考。（2）檢測老年病房ABHAP患者痰標本中分離的AB菌株在非生物體（abiotic）表面生物被膜（BF）形成能力，探討鮑曼不動桿菌生物被膜形成能力與所致醫(yī)院獲得性肺炎的相關性。（3）檢測臨床分離的AB菌株對肺上皮細胞的黏附性，探討鮑曼不動桿菌對生物體（biotic）的黏附能力與所致醫(yī)院獲得性肺炎的相關性。（4）檢測鮑曼不動桿菌生物被膜相關基因，分析影響生物被膜形成的內在因素。 2、方法：（1）制定病例調查表，，收集121例AB所致老年患者HAP病例的基本情況、既往應用抗生素的種類、基礎疾病、臨床表現(xiàn)、藥敏結果及治療轉歸等，尋找易感因素，明確所致感染的臨床特點及預后情況。（2）96孔聚苯乙烯板構建生物被膜模型，采用結晶紫染色法檢測AB-BF的形成能力，通過病例對照研究，比較不同生物被膜形成能力的AB菌株所致HAP臨床觀測指標及抗菌藥物耐藥性的差異。（3）采用吉姆薩染色法計算黏附于肺上皮細胞A549的細菌數(shù)目，以標準菌株ATCC19606T為對照，判斷鮑曼不動桿菌對生物體表面的黏附能力。（4）使用聚合酶鏈式反應（PCR）方法檢測鮑曼不動桿菌的生物被膜相關基因。 3、結果：（1）AB所致老年HAP患者伴隨基礎疾病較多，多有體內留置導管史，發(fā)病前多使用廣譜抗生素；57.9%無發(fā)熱，白細胞升高者僅占41.3%，而中性粒細胞百分比及C-反應蛋白（CRP）升高者分別占76.0%、81.8%；胸片多表現(xiàn)為斑片狀滲出影（94.2%）；多藥耐藥株及泛耐藥株分別占79.3%和24.0%；以30d為終點的logistic回歸分析顯示，APACHE II評分是ABHAP死亡的獨立危險因素。（2）分離自老年病房呼吸道的AB菌株均具有BF形成能力，其中27.3%表現(xiàn)為強陽性；隨著BF形成能力的增強，AB對慶大霉素、米諾環(huán)素、頭孢他啶的耐藥性顯著下降；分離自APACHE II評分較低或非ICU患者的菌株，生物被膜形成能力較強；不同BF形成能力的AB菌株所致HAP臨床表現(xiàn)及30d病死率差異無統(tǒng)計學意義。 （3）121株AB菌株對肺上皮細胞具有一定的黏附能力，其中強黏附能力菌株占33.9%；AB對A549細胞的黏附數(shù)目與體外BF形成能力呈正相關；強黏附能力組對大多數(shù)抗生素的耐藥性較弱黏附能力組顯著降低；不同黏附能力的AB菌株所致HAP的臨床表現(xiàn)及30d病死率差異無統(tǒng)計學意義。（4）生物被膜強陽性組的基因擴增陽性率均高于弱陽性組，其中兩組間csuAB、csuC及bfmS基因擴增率的差異有統(tǒng)計學意義。 結論：（1）AB所致老年患者HAP臨床表現(xiàn)多不典型，中性粒細胞比例及CRP升高具有較高敏感性，胸部X線多表現(xiàn)為支氣管肺炎的征象；老年患者痰標本分離的AB呈現(xiàn)多重耐藥及泛耐藥現(xiàn)象；APACHE II評分是老年患者ABHAP死亡的獨立危險因素。（2）隨著BF形成能力的增強，AB對某些抗生素耐藥性顯著下降；分離自APACHE II評分較低或非ICU患者的菌株，BF形成能力較強；不同BF形成能力的菌株對所致老年HAP的臨床表現(xiàn)及預后無影響。（3）AB對A549細胞的黏附數(shù)目與體外BF形成能力呈正相關；不同黏附能力的菌株對所致老年HAP的臨床表現(xiàn)及預后無顯著影響。（4）AB-BF形成是眾多因素調控的結果，BF相關基因csuA/BACDE、bap、abaI、bfmS廣泛存在于分離自下呼吸道的鮑曼不動桿菌臨床菌株中，強陽性組菌株的基因擴增率高于弱陽性組。
[Abstract]:1, objective: (1) retrospective analysis of the hospital acquired pneumonia (HAP) cases of elderly patients with Acinetobacter Bauman (AB) in our hospital for 2010-2012 years, and summarize the clinical characteristics, the susceptible factors and the resistance phenotype, and provide reference for the rational use of antibiotics in clinical. (2) the isolated AB strains isolated from the sputum specimens of ABHAP patients in the senile disease room were in non living organisms (abiotic). The formation ability of surface biofilm (BF) was used to investigate the correlation between the formation ability of Acinetobacter Bauman biofilm and hospital acquired pneumonia. (3) detection of the adhesion of clinical isolates of AB strains to lung epithelial cells, and the correlation between the adhesion ability of Acinetobacter sp. Bauman to the organism (biotic) and the result of hospital acquired pneumonia. (4) The genes related to biofilm of Acinetobacter baumannii were measured, and the internal factors affecting biofilm formation were analyzed.
2, methods: (1) to formulate a case questionnaire to collect the basic situation of HAP cases in 121 elderly patients with AB, and to use the types of antibiotics, basic diseases, clinical manifestations, drug sensitivity results and treatment outcomes, to find the susceptibility factors and to clarify the clinical characteristics and prognosis of the infection. (2) 96 hole polystyrene board is used to construct a biofilm model, The formation ability of AB-BF was detected by crystal violet staining. By case control study, the clinical observation index of HAP and the difference of antimicrobial resistance caused by AB strains with different biofilm formation ability were compared. (3) the number of bacteria adhered to A549 in lung epithelial cells was calculated by GIM SA staining method, and the standard strain ATCC19606T was used as the control. The adhesion ability of Acinetobacter baumannii to the surface of the organism was broken. (4) polymerase chain reaction (PCR) was used to detect the biofilm related genes of Acinetobacter baumannii from Bauman. Methods: the genes of Acinetobacter baumannii were detected by polymerase chain reaction (PCR).
3, the results were as follows: (1) the elderly patients with AB were associated with more basic diseases, most of which had a history of indwelling catheter in the body, and most of them used broad-spectrum antibiotics before the onset of the disease; 57.9% had no fever, only 41.3% of the white blood cells were elevated, while the percentage of neutrophils and the increase of C- reactive protein (CRP) were 76% and 81.8%, and the chest films showed plaque like exudative shadow (94.2%). The multidrug-resistant and pan drug resistant strains accounted for 79.3% and 24%, respectively. The logistic regression analysis at the end of 30d showed that APACHE II score was an independent risk factor for ABHAP death. (2) AB strains isolated from the respiratory tract of the elderly ward had BF formation ability, and 27.3% of them were strong Yang; with the enhancement of BF formation ability, AB against gentamicin, The drug resistance of minocycline and ceftazidime decreased significantly; the strains isolated from patients with low or non ICU APACHE II scores had stronger biofilm formation ability, and there was no significant difference in the clinical manifestation of HAP and the difference in the mortality rate of 30d caused by AB strains with different BF formation ability.
(3) 121 strains of AB strain had a certain adhesion to the lung epithelial cells, of which the strong adhesion ability was 33.9%; the adhesion number of AB to the A549 cells was positively correlated with the ability to form the BF in vitro; the strong adhesion ability group decreased the resistance of most antibiotics to the weak adhesion ability group; the AB strain of the different adhesion ability caused HAP. There was no significant difference in the clinical manifestation and the mortality of 30d. (4) the positive rate of gene amplification in the strong positive biofilm group was higher than that in the weak positive group, and the difference in the amplification rate of the csuAB, csuC and bfmS genes among the two groups was statistically significant.
Conclusions: (1) the clinical manifestations of HAP in the elderly patients with AB were mostly untypical, the proportion of neutrophils and the increase of CRP had high sensitivity, and the chest X ray showed the signs of bronchopneumonia, and the AB of the sputum specimens in the elderly showed multidrug resistance and pan resistance, and the evaluation of APACHE II was an independent risk factor for the death of ABHAP in the elderly patients. 2) with the enhancement of BF formation ability, the resistance of AB to some antibiotic resistance was significantly decreased; the strains isolated from patients with lower APACHE II score or non ICU were stronger, and the strains with different BF formation ability had no effect on the clinical manifestation and prognosis of the aged HAP. (3) the number of AB to A549 cells and the ability to form BF in vitro was positive. The strains with different adhesion ability have no significant effect on the clinical manifestation and prognosis of the elderly HAP. (4) the formation of AB-BF is the result of the regulation of many factors. The BF related genes csuA/BACDE, BAP, abaI, bfmS exist widely in the clinical strains isolated from the lower respiratory tract, and the gene amplification rate of the strong positive group is higher than that of the weak Yang. Sex group.
【學位授予單位】：中國人民解放軍醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2013
【分類號】：R563.1

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