本文選題：醫(yī)院感染 + 血流感染�。� 參考：《天津醫(yī)科大學(xué)》2007年碩士論文

【摘要】： 醫(yī)院感染發(fā)病率日益增加，危害嚴(yán)重。醫(yī)院感染的暴發(fā)，導(dǎo)致病死率高，且延長(zhǎng)住院時(shí)間，增加住院費(fèi)用，影響惡劣�？刂漆t(yī)院感染暴發(fā)的關(guān)鍵是如何早期發(fā)現(xiàn)，及時(shí)控制。表型分型方法包括生化分型、抗生素藥敏譜分型等方法，存在諸多不足，如分辨力低，影響因素多，重復(fù)性較差等。分子分型技術(shù)應(yīng)用于醫(yī)院感染暴發(fā)的流行病學(xué)調(diào)查有重要意義。 本研究對(duì)近期某院術(shù)后28株洋蔥伯克霍爾德菌醫(yī)院血流感染臨床株進(jìn)行表型和基因分型研究。探討應(yīng)用分子流行病方法調(diào)查洋蔥伯克霍爾德菌醫(yī)院血流感染暴發(fā)的價(jià)值。 第一部分洋蔥伯克霍爾德茵血流感染臨床株的抗生素藥敏譜分析 目的：了解28株洋蔥伯克霍爾德菌醫(yī)院血流感染臨床株的抗生素藥敏譜。 方法：應(yīng)用瓊脂紙片擴(kuò)散法(K-B法)和微量肉湯稀釋法(MICs)檢測(cè)28株臨床株對(duì)15種抗生素的抗生素藥敏譜。 結(jié)果：洋蔥伯克霍爾德菌醫(yī)院血流感染臨床株，對(duì)15種藥物敏感實(shí)驗(yàn)的結(jié)果表明，全部菌株對(duì)四環(huán)素、阿米卡星、慶大霉素耐藥，對(duì)氟喹諾酮、碳?xì)涿赶╊悺?fù)方新諾明、米諾環(huán)素、頭孢曲松、頭孢他啶、哌拉西林敏感，對(duì)頭孢哌酮舒巴坦鈉、頭孢吡肟、頭孢氨噻肟的敏感性存在微小差異。 第二部分洋蔥伯克霍爾德菌血流感染臨床株的recA基因分型研究 目的：28株洋蔥伯克霍爾德菌醫(yī)院血流感染臨床株的recA基因分型研究。 方法：進(jìn)行recA基因分型研究： 1．對(duì)28株洋蔥伯克霍爾德菌醫(yī)院血流感染臨床株進(jìn)行recA基因的PCR擴(kuò)增。 2．應(yīng)用限制性內(nèi)切酶HaeⅢ對(duì)recA基因PCR擴(kuò)增片段進(jìn)行PCR-RFLP分析。 3．recA基因PCR擴(kuò)增片段的序列分析。 結(jié)果：應(yīng)用PCR-RFLP技術(shù)對(duì)28株臨床株進(jìn)行recA基因分型，結(jié)果表明全部臨床株recA基因的PCR擴(kuò)增陽(yáng)性，PCR產(chǎn)物經(jīng)HaeⅢ限制性內(nèi)切酶反應(yīng)，出現(xiàn)一致的限制性內(nèi)切酶反應(yīng)譜型，符合RFLP-J型，即B．stabilis(基因型Ⅳ)。recA基因PCR擴(kuò)增片段的序列分析，進(jìn)一步證實(shí)為B．stabilis(基因型Ⅳ)。 第三部分洋蔥伯克霍爾德菌血流感染臨床株的脈沖場(chǎng)凝膠電泳(PFGE)分析 目的：了解應(yīng)用脈沖場(chǎng)凝膠電泳(PFGE)在洋蔥伯克霍爾德菌醫(yī)院血流感染暴發(fā)調(diào)查中的作用。 方法：從28株分離株中隨機(jī)選取一株未經(jīng)recA基因測(cè)序的樣本B140(抗生素藥敏譜型Ⅰ)和一株經(jīng)recA基因測(cè)序的樣本B258(抗生素藥敏譜型Ⅴ)，應(yīng)用限制性內(nèi)切酶SpeⅠ進(jìn)行全基因組DNA的脈沖場(chǎng)凝膠電泳(PFGE)分析。 結(jié)果：臨床株B140和B258全基因組DNA的限制性內(nèi)切酶反應(yīng)圖譜具有同樣的條帶數(shù)，且相應(yīng)條帶大小相同，依照Tenover等的標(biāo)準(zhǔn)，判斷為同一克隆。 結(jié)論： 1．本次研究的致病菌區(qū)別于區(qū)別于囊性纖維化(CF)患者的呼吸系統(tǒng)流行的多重耐藥菌株。依據(jù)對(duì)頭孢哌酮舒巴坦鈉、頭孢吡肟、頭孢氨噻肟的敏感性存在的微小差異，，28株臨床株的抗生素藥敏譜表現(xiàn)為七型。 2．應(yīng)用PCR-RFLP技術(shù)對(duì)28株臨床株進(jìn)行recA基因分型，結(jié)果表明全部臨床株recA基因的PCR擴(kuò)增陽(yáng)性，PCR產(chǎn)物經(jīng)HaeⅢ限制性內(nèi)切酶反應(yīng)，出現(xiàn)一致的限制性內(nèi)切酶反應(yīng)譜型，符合RFLP-J型，即B．stabilis(基因型Ⅳ)。recA基因PCR擴(kuò)增片段的序列分析，進(jìn)一步證實(shí)為B．stabilis(基因型Ⅳ)。 3．從分離株中，隨機(jī)選取一株未經(jīng)recA基因測(cè)序的樣本B140(抗生素藥敏譜型Ⅰ)和一株經(jīng)recA基因測(cè)序的樣本B258(抗生素藥敏譜型Ⅴ)，進(jìn)行脈沖場(chǎng)凝膠電泳(PFGE)分析，結(jié)果表明2株臨床株的全基因組DNA的限制性內(nèi)切酶SepⅠ反應(yīng)圖譜，有同樣的條帶數(shù)，且相應(yīng)條帶大小相同，依照Tenover等的標(biāo)準(zhǔn)，判斷2株臨床株為同一克隆。 4．綜上研究，經(jīng)抗生素藥敏譜分型、recA基因的PCR-RFLP分型和序列分析、以及脈沖場(chǎng)凝膠電泳分型研究，可以判定2005年6月至7月某醫(yī)院手術(shù)后發(fā)生的28例敗血癥患者為B．stabilis(基因型Ⅳ)的醫(yī)院血流感染暴發(fā)。 5．recA基因的PCR-RFLP分型和脈沖場(chǎng)凝膠電泳分型對(duì)洋蔥伯克霍爾德菌的分型比較，均顯示相同的分型結(jié)果，對(duì)醫(yī)院感染暴發(fā)的調(diào)查及追蹤溯源有重大價(jià)值。以經(jīng)濟(jì)和易操作性的角度，對(duì)洋蔥伯克霍爾德菌醫(yī)院感染暴發(fā)的調(diào)查，recA基因的PCR-RFLP分型更具推廣價(jià)值。 6．本研究醫(yī)院血流感染暴發(fā)中，洋蔥伯克霍爾德菌作為致病菌應(yīng)引起高度重視。
[Abstract]:The incidence of nosocomial infection is increasing and the harm is serious. The outbreak of nosocomial infection leads to high mortality, prolongs hospitalization time and increases hospitalization expenses. The key of controlling nosocomial infection outbreaks is how to discover early and control timely. The methods of phenotypic typing include biochemical typing, antibiotic susceptibility typing, and so on, there are many shortcomings. Such as low resolution, many influencing factors and poor repeatability, etc. molecular typing technology is of great significance in the epidemiological investigation of nosocomial infection outbreaks.
In this study, the phenotypic and genotyping of 28 clinical strains of Burke and Holder fungus nosocomial flow infection in a hospital of onions were studied in this study. The value of molecular epidemiological method was investigated to investigate the outbreak of blood flow infection in Burke and Holder fungus hospital.
Part 1 antibiotic susceptibility spectrum analysis of clinical isolates of Burke Holder blood stream infected with onion
Objective: to understand the antibiotic susceptibility spectrum of 28 strains of Burke Holder fungus hospital infection.
Methods: agar disk diffusion method (K-B) and micro broth dilution (MICs) were used to detect antibiotic susceptibility spectrum of 28 clinical isolates to 15 antibiotics.
Results: the clinical strain of the bloodstream infection of the onions Burke Holder hospital, the results of 15 drug sensitive experiments showed that all strains were resistant to tetracycline, Amikacin, gentamicin, fluoroquinolone, carbenolene, minocycline, ceftriaxone, ceftazidime, piperacillin, cefoperazone, and cefoperazine, and cefoperazone, sulbactam, and cefoperazone. The sensitivity of pyrazoxime and cefotaxime is slightly different.
The second part is recA genotyping of clinical isolates of Burke Holder infected blood stream in onion.
Objective: To study the recA genotyping of 28 strains of clinical isolates of Burke Holder disease in onion hospital.
Methods: the study of recA genotyping:
1. PCR amplification of recA gene was carried out on 28 strains of clinical isolates of Burke Holder fungus hospital infection.
2. restriction endonuclease Hae III was used to carry out PCR-RFLP analysis of recA gene PCR amplification fragment.
Sequence analysis of PCR amplified fragment of 3.recA gene.
Results: the recA genotyping of 28 strains of clinical strains was carried out by PCR-RFLP technique. The results showed that the PCR amplification of recA gene was positive in all clinical strains. The PCR product, through the Hae III restriction endonuclease reaction, appeared a consistent restriction endonuclease response spectrum type, which conformed to the RFLP-J type, that is, the sequence of B.stabilis (genotype IV).RecA gene PCR amplification fragment. It is further confirmed that B.stabilis (genotype IV).
The third part is the pulsed field gel electrophoresis (PFGE) analysis of clinical isolates of Burke Holder blood stream infection in onion.
Objective: To investigate the role of pulsed field gel electrophoresis (PFGE) in investigating the outbreak of bloodstream infection in Burke Holder fungus hospital.
Methods: a sample of B140 (antibiotic susceptibility type I) and a sample of B258 (antibiotic susceptibility type V), which were sequenced by recA gene, were randomly selected from 28 isolates, and the restriction endonuclease Spe I was used to analyze the whole genome DNA by pulse field gel electrophoresis (PFGE).
Results: the restriction endonuclease response map of B140 and B258 genome DNA has the same number of bands, and the corresponding band size is the same. According to the standard of Tenover and so on, it is judged to be the same clone.
Conclusion:
1. the pathogenic bacteria in this study were distinguished from the multidrug-resistant strains prevalent in the respiratory system of patients with cystic fibrosis (CF). According to the slight differences in the sensitivity of cefoperazone sulbactam, cefepime, cefotaxime, the antibiotic susceptibility of 28 clinical strains was shown to be type seven.
2. the recA genotyping was performed on 28 strains of clinical strains by PCR-RFLP technology. The results showed that the PCR amplification of the recA gene was positive in all clinical strains. The PCR product, through the Hae III restriction endonuclease reaction, appeared a consistent restriction endonuclease response spectrum type, which conformed to the sequence analysis of RFLP-J type, namely, B.stabilis (genotype IV).RecA gene PCR amplification fragment. One step was confirmed as B.stabilis (genotype IV).
3. from the isolates, a sample of B140 (antibiotic susceptibility type I) and a sample B258 (antibiotic susceptibility profile V), which were sequenced by recA gene, were randomly selected and analyzed by pulse field gel electrophoresis (PFGE). The results showed that the restriction endonuclease Sep I reaction Atlas of the total genomic DNA of the total genome of 2 strains of clinical strains was similar. The number of bands and the size of the corresponding bands were the same. According to the criteria such as Tenover, 2 clinical isolates were identified as the same clone.
4. in a comprehensive study, 28 cases of septicemia in a hospital from June 2005 to July were diagnosed as B.stabilis (genotype IV) in hospital blood flu outbreak, after the antibiotic susceptibility typing, PCR-RFLP typing and sequence analysis of recA gene, and pulse field gel electrophoresis typing.
The comparison of the typing of 5.recA gene PCR-RFLP and pulse field gel electrophoresis on the typing of Burke and Holder bacteria in onion showed the same typing results. It was of great value for the investigation and tracing of the nosocomial outbreak. The investigation of the outbreak of the nosocomial infection of the onion Burke and Holder bacteria, the P of the recA gene in the perspective of economic and easy operation The CR-RFLP classification is more valuable.
6. in the study of outbreak of nosocomial bloodstream infection, onion Burke Holder bacteria should be highly regarded as pathogenic bacteria.

【學(xué)位授予單位】：天津醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2007
【分類號(hào)】：R181.3

【引證文獻(xiàn)】

相關(guān)碩士學(xué)位論文 前1條

1 肖丹;射陽(yáng)鹽場(chǎng)地區(qū)鯽源嗜水氣單胞菌分子流行病學(xué)研究[D];上海海洋大學(xué);2011年

本文編號(hào)：1804049