本文關(guān)鍵詞：碳青霉烯類耐藥肺炎克雷伯菌耐藥機(jī)制及同源性研究　出處：《浙江大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 碳青霉烯類 肺炎克雷伯菌 耐藥 碳青霉烯酶 脈沖場(chǎng)凝膠電泳 多位點(diǎn)序列分型 全基因組測(cè)序 同源性

【摘要】：肺炎克雷伯菌是院內(nèi)獲得性感染中最為常見的致病菌之一,主要引起肺炎、敗血癥、尿路感染、肝膿腫及腹膜炎等。隨著抗菌藥物的廣泛使用,尤其是碳青霉烯類抗生素的大量或不合理使用,碳青霉烯類耐藥肺炎克雷伯菌(Carbapenem-resistant Klebsiella pneumonia, CRKP)呈現(xiàn)全球播散趨勢(shì),在我國(guó)分離率逐年攀升,對(duì)臨床治療帶來極大挑戰(zhàn)。本研究旨在探討本院CRKP院內(nèi)感染相關(guān)危險(xiǎn)因素和耐藥機(jī)制,并分析其同源性和院內(nèi)感染流行現(xiàn)狀,為CR KP的防控提供科學(xué)的理論依據(jù)。 本研究收集了來自本院8個(gè)臨床科室的30株CRKP,制定統(tǒng)一調(diào)查表收集臨床資料并進(jìn)行分析。采用紙片擴(kuò)散法進(jìn)行常見藥物的體外敏感性實(shí)驗(yàn),并用改良Hodge試驗(yàn)檢測(cè)所有菌株是否產(chǎn)碳青霉烯酶。所有菌株均利用脈沖場(chǎng)凝膠電泳(Pulsed field gel electrophoresis, PFGE)和多位點(diǎn)序列分型(Multilocus sequencetyping,MLST)兩種技術(shù)進(jìn)行同源性分析,部分菌株進(jìn)行全基因組測(cè)序(Whole genome sequencing, WGS)分析親緣關(guān)系遠(yuǎn)近。 臨床資料顯示30位感染患者的平均年齡較高(72±15歲),各項(xiàng)病情評(píng)分指標(biāo)均顯示病情嚴(yán)重�；A(chǔ)疾病中神經(jīng)系統(tǒng)疾病、糖尿病和心力衰竭患者占了前三位。而有90%的患者在三個(gè)月內(nèi)使用過p-內(nèi)酰胺酶抑制劑合劑,50%多的患者使用過碳青霉烯類。近三個(gè)月內(nèi)絕大多數(shù)患者曾留置導(dǎo)管,例如導(dǎo)尿管、深靜脈導(dǎo)管和氣管導(dǎo)管等。體外藥物敏感性實(shí)驗(yàn)顯示所有菌株對(duì)美羅培南和厄他培南耐藥,除一株對(duì)亞胺培南為中介外其余也全部耐藥。對(duì)三、四代頭孢、復(fù)合制劑所有菌株均耐藥。而對(duì)環(huán)丙沙星、慶大霉素和阿米卡星的耐藥率分別是90.0%、97.0%和87.0%。所有菌株對(duì)多粘菌素E和替加環(huán)素均無耐藥。耐藥酶基因檢測(cè)結(jié)果提示30株肺炎克雷伯菌在攜帶blaKPC-2的同時(shí)均帶有blasHV。此外,有部分菌株還同時(shí)帶有blacTX-M和blaTEM。PFGE將所有菌株分為A-H8個(gè)克隆群,主要有2個(gè)克隆群出現(xiàn)院內(nèi)播散,分別是A和C克隆群。MLST分析得到5種序列型(Sequence type, ST),其中23株屬于ST11,3株屬于ST437,2株屬于ST290,另外兩株分別為ST133和ST15。5株A克隆(ST11)菌株的全基因組測(cè)序?qū)?株細(xì)菌分為3個(gè)不同的群組(2株：2株：1株),同一群組的菌株為一簇,親緣關(guān)系密切,單獨(dú)的那株自成一簇,與前兩簇關(guān)系疏遠(yuǎn)。因此,WGS較PFGE和MLST更深入的揭示了菌株的遺傳背景,更利于菌株的溯源追蹤。 綜上所述,CRKP感染患者具有多種危險(xiǎn)因素,患者預(yù)后情況與多種危險(xiǎn)因素和治療相關(guān)。初始經(jīng)驗(yàn)性抗感染治療應(yīng)根據(jù)感染危險(xiǎn)因素進(jìn)行選擇,并及時(shí)綜合分析患者病情,結(jié)合藥敏結(jié)果進(jìn)行目標(biāo)治療。產(chǎn)肺炎克雷伯菌碳青霉烯酶(Klebsiella pneumoniae carbapenemase, KPC)2型是本院CRKP耐藥的重要原因,多種耐藥基因的共同攜帶導(dǎo)致其多重耐藥表型。ST11型克隆播散是本院感染流行的重要原因,應(yīng)加強(qiáng)醫(yī)院感染監(jiān)控力度。
[Abstract]:Klebsiella pneumoniae is one of the most common pathogens in nosocomial infection, which mainly causes pneumonia, sepsis, urinary tract infection, liver abscess and peritonitis. In particular, a large number of carbapenem antibiotics or unreasonable use. Carbapenem resistant Klebsiella pneumonia. CRKP) presents a global spread trend, and the isolation rate in China is increasing year by year, which brings great challenge to clinical treatment. This study aims to explore the risk factors and drug resistance mechanism of hospital CRKP nosocomial infection. The homology of CR KP and the prevalence of nosocomial infection were analyzed in order to provide a scientific theoretical basis for the prevention and control of CR KP. In this study, 30 CRKPs from 8 clinical departments were collected, and a unified questionnaire was developed to collect and analyze the clinical data. In vitro sensitivity tests of common drugs were carried out by disk diffusion method. The improved Hodge test was used to detect whether all strains produced carbapenem. All strains were detected by pulsed field gel electrophoresis (PGE). Pulsed field gel electrophoresis. PFGE and multilocus sequencing typing (MLST) were used for homology analysis. Some strains were analyzed by whole genome sequencing (WGS). Clinical data showed that the average age of 30 infected patients was higher than 72 鹵15 years old. Diabetes and heart failure accounted for the top three. 90% of the patients had used plactamase inhibitors within three months. More than 50% patients have used carbapenems. The vast majority of patients have had catheters, such as catheters, in the last three months. In vitro drug sensitivity tests showed that all the strains were resistant to meropenem and ertapenem, except one for imipenem. All strains of compound preparation were resistant to ciprofloxacin, gentamicin and amikacin, respectively. The resistance rates of ciprofloxacin, gentamicin and amikacin were 90.0%. 97.0% and 87.0. All strains were not resistant to polymyxin E and tegacyclin. The results of enzyme resistance gene analysis showed that all 30 strains of Klebsiella pneumoniae carried blaKPC-2 with b. LasHV. in addition. Some strains also carry blacTX-M and blaTEM.PFGE to divide all strains into A-H 8 clone groups, mainly 2 clones spread in the hospital. According to the analysis of A and C clones. MLST, 5 kinds of sequence type and STN were obtained, of which 23 belonged to ST11N, 3 of which belonged to ST437. Two strains belong to ST290. The other two strains, ST133 and ST15.5 strain A clone ST11, were sequenced and divided into three different groups: 2 strains: 2 strains: 1 strain). The strains of the same group are a cluster, closely related to each other, and the single strain is isolated from the first two clusters. Therefore, the genetic background of the strain is revealed more deeply by WGS than by PFGE and MLST. More conducive to traceability of the strain tracking. In conclusion, there are many risk factors in patients with CRKP infection, and the prognosis of patients is related to many risk factors and treatment. The initial empirical anti-infection therapy should be based on the risk factors of infection. And timely comprehensive analysis of the patient's condition. Combined with the results of drug sensitivity, Klebsiella pneumoniae carbapenemase was produced by Klebsiella pneumoniae carbapenemase, Klebsiella pneumoniae carbapenemase was produced by Klebsiella pneumoniae. KPC)2 type is an important cause of CRKP resistance in our hospital. The common carrying of multidrug resistance genes leads to the spread of multidrug resistance phenotype. ST11 is an important reason for the prevalence of infection in our hospital. Hospital infection monitoring should be strengthened.
【學(xué)位授予單位】：浙江大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R446.5

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本文編號(hào)：1404329