【摘要】：屎腸球菌(Enterococcus faecium)屬于革蘭氏陽(yáng)性、過(guò)氧化氫酶陰性球菌,是人和動(dòng)物腸道內(nèi)的主要菌群。自上世紀(jì)90年代以來(lái),屎腸球菌逐漸成為人和動(dòng)物重要的條件性致病菌。在我國(guó),對(duì)屎腸球菌的研究主要集中于人感染屎腸球菌的病例報(bào)道,及分離菌株的分類研究,在動(dòng)物方面,則少有報(bào)道。本研究成功分離到18株腸球菌臨床分離株,對(duì)進(jìn)一步研究腸球菌診斷方法,治療腸球菌感染以及耐藥機(jī)制和多位點(diǎn)序列分型提供基礎(chǔ)。首先通過(guò)細(xì)菌分離,染色鑒定,凝固酶試驗(yàn),延伸因子tuf及16S rRNA序列鑒定,從采自甘肅、青海等地的羊及豬組織病料中,分離到18株屎腸球菌,分別命名為L(zhǎng)VRI1101,LVRI1102,LVRI1103,LVRI1104,LVRI1301,LVRI1302,LVRI1303,LVRI1304,YYAN1401,YYAN1402,YYAN1403,YYAN1404,lby1,lby2,lby3,lbg1,lbg2,lbg3。藥敏試驗(yàn)結(jié)果表明,所分離菌株均為多重耐藥性菌株。小鼠致病性實(shí)驗(yàn)結(jié)果表明,所分離菌株均具有致病力。多位點(diǎn)序列分型(multilocus sequence type,MLST)結(jié)果發(fā)現(xiàn),所分離菌株屬于3個(gè)新的序列型(Sequence type),分別命名為Enterococcus faecium ST989,Enterococcus faecium ST1017,Enterococcus faecium ST1018,已添加到數(shù)據(jù)庫(kù)中。由于目前在基層尚缺乏快速有效的屎腸球菌血清學(xué)診斷方法,從而可能會(huì)造成誤診及漏診,因此,我們采用超聲細(xì)胞破碎法提取屎腸球菌菌體抗原作為抗原,致敏醛化鞣酸化的綿陽(yáng)紅細(xì)胞,與屎腸球菌高免兔血清,屎腸球菌陽(yáng)性血清進(jìn)行間接血凝實(shí)驗(yàn)。結(jié)果顯示該方法特異性好、敏感性高,建立了檢測(cè)屎腸球菌抗體的間接血凝實(shí)驗(yàn)方法(IHA),可為屎腸球菌病的血清學(xué)快速診斷及流行病學(xué)調(diào)查提供依據(jù)。
[Abstract]:Enterococcus faecium (Enterococcus faecium) belongs to Gram-positive and catalase negative cocci. Since 1990 s, Enterococcus faecium has become an important conditional pathogen in human and animal. In China, the studies on Enterococcus faecium mainly focus on the case reports of Enterococcus faecium infection in human beings, and the taxonomic studies of isolated strains, but there are few reports on animals. In this study, 18 strains of Enterococcus were successfully isolated, which provided the basis for further study on the diagnosis of Enterococcus, the treatment of enterococcal infection, the mechanism of drug resistance and the classification of multilocus sequence. At first, 18 strains of Enterococcus faecium were isolated from sheep and pig tissue samples collected from Gansu, Qinghai and other places by bacterial isolation, staining, coagulase test, extension factor tuf and 16s rRNA sequence identification. They were named LVRI1101,LVRI1102,LVRI1103,LVRI1104, respectively. LVRI1301,LVRI1302,LVRI1303,LVRI1304,YYAN1401,YYAN1402,YYAN1403,YYAN1404,lby1,lby2,lby3,lbg1,lbg2,lbg3. The results of drug sensitivity test showed that the isolates were multidrug resistant. The pathogenicity test of mice showed that all the isolated strains had pathogenicity. The results of multi-locus sequence typing (multilocus sequence type,MLST) showed that the isolated strains belonged to three new sequence-type (Sequence type), named Enterococcus faecium ST989,Enterococcus faecium ST1017,Enterococcus faecium ST1018, which had been added to the database. Due to the lack of a rapid and effective serological diagnosis method for Enterococcus faecium at present, it may lead to misdiagnosis and missed diagnosis. Therefore, we use the method of ultrasonic cell fragmentation to extract Enterococcus faecium antigen as antigen. The indirect hemagglutination test was carried out between the sensitized mianyang erythrocytes and the high immune rabbit serum of Enterococcus faecium and the positive serum of Enterococcus faecium. The results showed that the method had good specificity and high sensitivity. The indirect hemagglutination assay (IHA),) for the detection of Enterococcus faecium antibody could provide the basis for rapid serological diagnosis and epidemiological investigation of Enterococcus faecium.
【學(xué)位授予單位】：西北師范大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R446

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