本文選題：胥伐成格隆沙門(mén)菌　切入點(diǎn)：微生物敏感性試驗(yàn)　出處：《天津醫(yī)科大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：目的研究臨床分離胥伐成格隆沙門(mén)菌的抗菌藥物敏感性、分子分型、毒力基因攜帶和患者臨床特征,為了解胥伐成格隆沙門(mén)菌病原學(xué)特點(diǎn)、預(yù)防與控制其感染以及臨床合理使用抗菌藥物提供依據(jù)。方法1.收集2014年5月至10月我院腸道門(mén)診急性腹瀉患者的臨床資料,采集患者糞便標(biāo)本進(jìn)行沙門(mén)菌分離培養(yǎng)、生化鑒定、血清型分型。2.對(duì)鑒定得到的胥伐成格隆沙門(mén)菌采用K-B法和MIC法進(jìn)行11種抗菌藥物敏感性檢測(cè)。3.多位點(diǎn)序列分型(MLST):擴(kuò)增胥伐成格隆沙門(mén)菌的7個(gè)管家基因(thrA,purE,sucA,hisD,aroC,hemD,dnaN),將測(cè)序結(jié)果與國(guó)際數(shù)據(jù)庫(kù)網(wǎng)站比較后獲得等位基因圖譜和ST型。4.脈沖場(chǎng)凝膠電泳分型(PFGE):對(duì)胥伐成格隆沙門(mén)菌基因組DNA采用限制性內(nèi)切酶XbaI酶切,經(jīng)過(guò)脈沖場(chǎng)凝膠電泳后采用BioNumerics軟件對(duì)電泳條帶進(jìn)行聚類分析。5.分析患者病史和臨床表現(xiàn)以及與菌株分子分型的關(guān)系。6.毒力基因PCR檢測(cè):以沙門(mén)菌毒力基因島(SPI)1~5中的代表性基因(SPI1:invA、sitC、hilA;SPI2:sseL、sifA;SPI3:mgtC;SPI4:siiE;SPI5:sopB)、毒力質(zhì)�；�(pefA、prot6E、spvB)、調(diào)控基因(phoP)為目的基因,對(duì)菌株進(jìn)行特異引物PCR擴(kuò)增。結(jié)果1.共分離到胥伐成格隆沙門(mén)菌16株(14.8%),占同期全部非傷寒沙門(mén)菌(108株)的14.8%。2.16株胥伐成格隆沙門(mén)菌對(duì)11種抗菌藥物氨芐西林、氯霉素、鏈霉素、復(fù)方新諾明、四環(huán)素、萘啶酸、環(huán)丙沙星、左氧氟沙星、頭孢曲松、頭孢他啶、厄他培南均100%敏感。3.16株胥伐成格隆沙門(mén)菌的7個(gè)管家基因等位基因譜完全一致,依次是43、47、49、16、41、15、3,均為ST241型。4.以相似度100%為標(biāo)準(zhǔn),16株胥伐成格隆沙門(mén)菌共獲得3種PFGE帶型,包括A型14株、B型1株和C型1株。A型與B型相似度為94.2%,二者與C型相似度為77.8%。5.16例胥伐成格隆沙門(mén)菌感染患者男性10例、女性6例,中位年齡35.5歲,均因急性腹瀉就診,臨床診斷包括細(xì)菌性痢疾3例和胃腸炎13例,其中1例患者伴有高熱。有8例患者共同進(jìn)餐后發(fā)病,就診時(shí)間集中于2014年6月17日至19日的兩天之內(nèi),患者糞便分離菌株P(guān)FGE帶型完全一致,均為A型。6.16株胥伐成格隆沙門(mén)菌SPI1~5代表性基因(invA、sitC、hilA、sseL、sifA、mgtC、siiE、sopB)和調(diào)控基因(phoP)100%陽(yáng)性,毒力質(zhì)�；�(pefA、prot6E、spvB)均為陰性。結(jié)論1.2014年本地區(qū)出現(xiàn)多例胥伐成格隆沙門(mén)菌臨床感染,其中包含了一起感染暴發(fā)。2.本研究檢測(cè)到多種PFGE帶型ST241型胥伐成格隆沙門(mén)菌臨床株,對(duì)常用抗菌藥物尚保持敏感,但攜帶SPI1~5毒力基因和調(diào)節(jié)基因,提示其具有高致病力,對(duì)公眾健康構(gòu)成威脅。3.應(yīng)關(guān)注胥伐成格隆沙門(mén)菌臨床感染在我國(guó)的變化趨勢(shì)和危害,開(kāi)展持續(xù)監(jiān)測(cè)和研究。
[Abstract]:Objective to study the antimicrobial susceptibility, molecular typing, virulence gene carrying and clinical characteristics of Scheffarn's Salmonella schevaronii in order to understand its etiological characteristics. Prevention and control of infection and rational use of antimicrobial agents. Methods 1. Clinical data of patients with acute diarrhea in intestinal outpatient department from May 2014 to October were collected and stool samples were collected for isolation and culture of Salmonella. Biochemical identification, Serotype typing .2. using K-B method and MIC method to detect the susceptibility of 11 antimicrobial agents to the identified Salmonella schevaronii. Multilocus sequence typing: amplification of seven housekeeping genes of Schuffat into Salmonella grossurum DaroChemDdna nna, and sequencing. Results compared with the international database website, allelic map and St type. 4. Pulsed field gel electrophoresis (PGE) typing: the genomic DNA of Salmonella schevatensis was digested by restriction endonuclease XbaI. After pulse field gel electrophoresis, BioNumerics software was used to cluster analysis of the electrophoresis bands. 5. Analysis of patient's history, clinical manifestation and relationship with strain molecular typing. 6. Detection of virulence gene PCR: in Salmonella virulence gene island. The representative gene SPI1: invAt sitCor hila, SPI2: sseLLnsifAn SPI3: MGtCn SPI4: siiEn SPI5: sopB, virulent plasmid gene pefAbuprot6EspvB, regulatory gene phoP, is the target gene. Results 1. A total of 16 strains of Salmonella vulgaris, which accounted for 108 strains of non-typhoid salmonella, were isolated from the bacteria by PCR. 2. 16 strains of Salmonella xulfengensis were isolated for 11 antimicrobial agents ampicillin, chloramphenicol, streptomycin. The alleles of seven housekeeping alleles of compound sulfamine, tetracycline, naphthalic acid, ciprofloxacin, levofloxacin, ceftriaxone, ceftazidime and ertapenem were identical in alleles of 7 alleles of Salmonella xuvalensis. The sequence was 43, 47, 49, 41C153, all of which were ST241 type. 4. According to similarity degree 100%, 16 strains of Salmonella xufangchengensis were found to have three PFGE banding patterns. The similarity between type A and type C was 94.2, and the similarity between them was 77.8.5.16 cases of schistosomiasis infection were male 10 cases, female 6 cases, median age 35.5 years old, all of them were treated with acute diarrhea, the average age was 35.5 years old, the average age was 35.5 years old, and the similarity between the two strains was 77.8% and 5.16 cases respectively, including 10 males and 6 females, with a median age of 35.5 years, and a median age of 35.5 years. Clinical diagnosis included 3 cases of bacillary dysentery and 13 cases of gastroenteritis, of which 1 case was accompanied by high fever. The PFGE banding patterns of fecal isolates from patients were identical and were all positive for type A. 6.16 SPI1~5 representative genes of Salmonella schevaronii, AhilsseSse, sifsif, mgtCsii, sopB, and the regulatory gene, phopt1, 100%, respectively, and the positive rate was 100%, and the positive rate was 100% (P < 0.05), and the positive rate was 100% (P < 0.05). The virulence plasmid gene was negative. Conclusion 1. There were many clinical infections of Salmonella schevarinensis in 2014, including an outbreak of infection. In this study, a variety of clinical strains of ST241 type of PFGE were detected, and the results showed that there were many clinical strains of Schulonella schevaronii in 2014, and the results were as follows: (1) in this study, there were many clinical infections of Salmonella schevaronii in this area, including an outbreak of infection. SPI1~5 virulence gene and regulatory gene are still sensitive to common antimicrobial agents, which indicates that it is highly virulent and poses a threat to public health .3.We should pay attention to the trend and harm of clinical infection of Salmonella xuvalchengensis in China. Carry out continuous monitoring and research.
【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】：2017
【分類號(hào)】：R446.5;R516.3

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