鲍曼不动杆菌生物被膜形成能力与所致老年患者医院获得性肺炎的关系

发布时间：2018-07-09 15:31

本文选题：鲍曼不动杆菌 + 生物被膜　；参考：《中国人民解放军医学院》2013年硕士论文

【摘要】：1、目的：（1）回顾性分析我院2010-2012年鲍曼不动杆菌（AB）所致老年患者医院获得性肺炎（HAP）病例，总结临床特点、易患因素及耐药表型，为临床合理应用抗菌药物提供参考。（2）检测老年病房ABHAP患者痰标本中分离的AB菌株在非生物体（abiotic）表面生物被膜（BF）形成能力，探讨鲍曼不动杆菌生物被膜形成能力与所致医院获得性肺炎的相关性。（3）检测临床分离的AB菌株对肺上皮细胞的黏附性，探讨鲍曼不动杆菌对生物体（biotic）的黏附能力与所致医院获得性肺炎的相关性。（4）检测鲍曼不动杆菌生物被膜相关基因，分析影响生物被膜形成的内在因素。 2、方法：（1）制定病例调查表，，收集121例AB所致老年患者HAP病例的基本情况、既往应用抗生素的种类、基础疾病、临床表现、药敏结果及治疗转归等，寻找易感因素，明确所致感染的临床特点及预后情况。（2）96孔聚苯乙烯板构建生物被膜模型，采用结晶紫染色法检测AB-BF的形成能力，通过病例对照研究，比较不同生物被膜形成能力的AB菌株所致HAP临床观测指标及抗菌药物耐药性的差异。（3）采用吉姆萨染色法计算黏附于肺上皮细胞A549的细菌数目，以标准菌株ATCC19606T为对照，判断鲍曼不动杆菌对生物体表面的黏附能力。（4）使用聚合酶链式反应（PCR）方法检测鲍曼不动杆菌的生物被膜相关基因。 3、结果：（1）AB所致老年HAP患者伴随基础疾病较多，多有体内留置导管史，发病前多使用广谱抗生素；57.9%无发热，白细胞升高者仅占41.3%，而中性粒细胞百分比及C-反应蛋白（CRP）升高者分别占76.0%、81.8%；胸片多表现为斑片状渗出影（94.2%）；多药耐药株及泛耐药株分别占79.3%和24.0%；以30d为终点的logistic回归分析显示，APACHE II评分是ABHAP死亡的独立危险因素。（2）分离自老年病房呼吸道的AB菌株均具有BF形成能力，其中27.3%表现为强阳性；随着BF形成能力的增强，AB对庆大霉素、米诺环素、头孢他啶的耐药性显著下降；分离自APACHE II评分较低或非ICU患者的菌株，生物被膜形成能力较强；不同BF形成能力的AB菌株所致HAP临床表现及30d病死率差异无统计学意义。（3）121株AB菌株对肺上皮细胞具有一定的黏附能力，其中强黏附能力菌株占33.9%；AB对A549细胞的黏附数目与体外BF形成能力呈正相关；强黏附能力组对大多数抗生素的耐药性较弱黏附能力组显著降低；不同黏附能力的AB菌株所致HAP的临床表现及30d病死率差异无统计学意义。（4）生物被膜强阳性组的基因扩增阳性率均高于弱阳性组，其中两组间csuAB、csuC及bfmS基因扩增率的差异有统计学意义。结论：（1）AB所致老年患者HAP临床表现多不典型，中性粒细胞比例及CRP升高具有较高敏感性，胸部X线多表现为支气管肺炎的征象；老年患者痰标本分离的AB呈现多重耐药及泛耐药现象；APACHE II评分是老年患者ABHAP死亡的独立危险因素。（2）随着BF形成能力的增强，AB对某些抗生素耐药性显著下降；分离自APACHE II评分较低或非ICU患者的菌株，BF形成能力较强；不同BF形成能力的菌株对所致老年HAP的临床表现及预后无影响。（3）AB对A549细胞的黏附数目与体外BF形成能力呈正相关；不同黏附能力的菌株对所致老年HAP的临床表现及预后无显著影响。（4）AB-BF形成是众多因素调控的结果，BF相关基因csuA/BACDE、bap、abaI、bfmS广泛存在于分离自下呼吸道的鲍曼不动杆菌临床菌株中，强阳性组菌株的基因扩增率高于弱阳性组。
[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.
【学位授予单位】：中国人民解放军医学院
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R563.1

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