本文關(guān)鍵詞： 公共場(chǎng)所 葡萄球菌 藥敏試驗(yàn) 分子流行病學(xué)　出處：《廣東藥學(xué)院》2015年碩士論文　論文類型：學(xué)位論文

【摘要】：目的調(diào)查廣州部分公共場(chǎng)所環(huán)境物體表面葡萄球菌污染情況,揭示其耐藥譜及分子特征,為預(yù)防和控制公共場(chǎng)所葡萄球菌污染提供科學(xué)依據(jù)。方法采用橫斷面研究設(shè)計(jì),調(diào)查地鐵、火車站、汽車站和醫(yī)院候診大廳葡萄球菌污染狀況;應(yīng)用一系列標(biāo)準(zhǔn)的微生物實(shí)驗(yàn)方法進(jìn)行菌株的分離和鑒定;采用紙片擴(kuò)散法對(duì)所有葡萄球菌進(jìn)行藥物敏感試驗(yàn);使用PCR技術(shù)檢測(cè)金黃色葡萄球菌中PVL基因和qac基因攜帶情況,并對(duì)金黃色葡萄球菌進(jìn)行多位點(diǎn)測(cè)序分型(multilocus sequencing typing,MLST);應(yīng)用多重PCR技術(shù)對(duì)耐甲氧西林金黃色葡萄球菌(methicillin-resistant Staphylococcus aureus,MRSA)的進(jìn)行SCC mec分型。結(jié)果葡萄球菌污染情況:共采集公共場(chǎng)所物體表面樣本1100份。葡萄球菌檢出率為59.64%(656/1100):金黃色葡萄球菌污染率為7.55%(83/1100),其中MRSA污染率為1.73%(19/1100);凝固酶陰性葡萄球菌污染率為52.09%(573/1100),其中耐甲氧西林凝固酶陰性葡萄球菌污染率為6.09%(67/1100)。各個(gè)場(chǎng)所葡萄球菌污染率由高到低依次為:地鐵(75.63%,242/320)醫(yī)院(66.25%,265/400)火車站(43.89%,79/180)汽車客運(yùn)站(35.00%,70/200);金黃色葡萄球菌污染率由高到低依次為:地鐵(11.25%,36/320)汽車客運(yùn)站(8.50%,17/200)火車站(6.67%,12/180)醫(yī)院(4.50%,18/400)。藥物敏感試驗(yàn):葡萄球菌對(duì)11種藥物均有不同程度耐藥,藥物按照耐藥率由高到低依次為:青霉素(94.05%)紅霉素(84.15%)克林霉素(57.62%)利福平(48.78%)甲氧芐啶(42.23%)慶大霉素(31.25%)莫西沙星(21.29%)妥布霉素(14.33%)頭孢西丁(12.65%)替考拉寧(3.66%)利奈唑胺(2.44%)。金黃色葡萄球菌對(duì)頭孢西丁和妥布霉素的耐藥率高于凝固酶陰性葡萄球菌,對(duì)克林霉素和紅霉素的耐藥率低于凝固酶陰性葡萄球菌,差異有統(tǒng)計(jì)學(xué)意義(P0.05),其他藥物耐藥率在兩組間無統(tǒng)計(jì)學(xué)差異;甲氧西林耐藥菌株和甲氧西林敏感菌株對(duì)紅霉素耐藥率無統(tǒng)計(jì)學(xué)差異,甲氧西林耐藥菌株對(duì)其他藥物的耐藥率均高于甲氧西林敏感菌株,差異有統(tǒng)計(jì)學(xué)意義(P0.05);各采樣場(chǎng)所間耐藥率存在一定差異。PVL基因和qac基因檢測(cè):有2株MRSA(2/19,10.53%)和9株甲氧西林敏感的金黃色葡萄球菌(methicillin-susceptible Staphylococcus aureus,MSSA)(14.06%,9/64)攜帶PVL基因,兩組間PVL基因攜帶率沒有統(tǒng)計(jì)學(xué)差異(Fisher確切概率法,P=1.000);有9株MRSA(47.37%,9/19)和0株MSSA(0.00%,0/64)攜帶qac基因,兩組間qac基因攜帶率有統(tǒng)計(jì)學(xué)差異(Fisher確切概率法,P0.001)。SCCmec分型:在19株MRSA中,SCCmecⅠ型有1株(5.26%),Ⅱ型有3株(15.79%),Ⅲ型有2株(10.53%),Ⅳa型有5株(26.32%)未分型的有8株(42.11%),未檢測(cè)到Ⅳb型、Ⅳc型、Ⅳd型和Ⅴ型。MLST:金黃色葡萄球菌共檢測(cè)到27種ST型別,頻率由高到低依次為:ST188(26.55%)ST5(17.19%)ST72(15.63%)ST398(12.50%)ST6、ST15和ST1462(6.25%)ST125(4.69%)ST59、ST97和ST2140(3.13%)ST8、ST25、ST30、ST88、ST221、ST338、ST537、ST789、ST808、ST1141、ST1311、ST1507、ST1860、ST2114、ST2605和ST2668(1.56%)。根據(jù)ST型別間聚類分析,共形成3個(gè)克隆群(CC5,CC59和CC72),4個(gè)成對(duì)體(D6,D30,D188和D398)和8個(gè)單體(ST8,ST15,ST25,ST88,ST97,ST789,ST1141和ST2668)。通過對(duì)MRSA菌株進(jìn)行菌株間樹狀聚類分析,19株MRSA菌株聚成5個(gè)型別,提示可能存在不同公共場(chǎng)所間菌株交叉?zhèn)鞑サ娘L(fēng)險(xiǎn)。結(jié)論廣州公共場(chǎng)所環(huán)境物體表面葡萄球菌污染情況嚴(yán)重;這些葡萄球菌對(duì)常用抗生素耐藥率較高;金黃色葡萄球菌中存在一定數(shù)量PVL基因陽性菌株,MRSA菌株中qac基因攜帶率較高;MRSA菌株出現(xiàn)醫(yī)院和社區(qū)來源并存的現(xiàn)象;ST型別與我國(guó)近期流行菌株相符,大部分菌株屬于國(guó)際流行菌株,MRSA可能存在不同場(chǎng)所間菌株交叉?zhèn)鞑サ娘L(fēng)險(xiǎn)。
[Abstract]:Objective to investigate the Guangzhou public environmental surface Staphylococcus contamination, revealing the resistance spectrum and molecular characteristics, and provide scientific basis for the prevention and control of public places of Staphylococcus contamination. Methods the cross-sectional design and investigation of the subway, train station, bus station and hospital waiting hall Staphylococcus contamination; separation and identification of strains microbial experimental method of applying a series of standard; disk diffusion method was used for drug sensitivity test of Staphylococcus aureus; with detection of Staphylococcus aureus using PCR PVL gene and QAC gene, and Staphylococcus aureus were characterized by multilocus sequence typing (multilocus sequencing, typing, MLST); the application of multiplex PCR methicillin resistant Staphylococcus aureus (methicillin-resistant Staphylococcus, aureus, MRSA) of SCC MEC. The results of grape Aureus contamination: public places were collected 1100 samples. The surface of Staphylococcus aureus detection rate was 59.64% (656/1100): Staphylococcus aureus contamination rate was 7.55% (83/1100), in which MRSA pollution rate was 1.73% (19/1100); coagulase negative Staphylococcus contamination rate was 52.09% (573/1100), including methicillin resistant coagulase negative Staphylococcus contamination rate was 6.09% (67/1100) in various places. Staphylococcus infection rate from high to low in turn: the subway (75.63%, 242/320) hospital (66.25%, 265/400) train station (43.89%, 79/180) car passenger transport stations (35%, 70/200); Staphylococcus aureus contamination rate from high to low the subway: (11.25%, 36/320) bus station (8.50%, 17/200) train station (6.67%, 12/180) hospital (4.50%, 18/400). Drug sensitivity test: Staphylococcus to 11 kinds of drugs have different degrees of resistance, according to the drug resistance rate from high to low: penicillin (94. 05% (84.15%)) erythromycin clindamycin (57.62%) rifampin (48.78%) and (42.23%) gentamicin (31.25%) moxifloxacin (21.29%) tobramycin (14.33%) cefoxitin (12.65%) and Ti Koa Laing (3.66%) linezolid (2.44%). Staphylococcus aureus resistant to cefoxitin and tobramycin was higher than that of coagulase negative Staphylococcus resistance to clindamycin and erythromycin was lower than that of coagulase negative staphylococcus, the difference was statistically significant (P0.05), other drug resistance rate showed no significant difference between the two groups; methicillin-resistant strains of methicillin and methicillin sensitive strains had no significant difference on erythromycin resistant strains of methicillin resistant, resistant to other drugs were higher than methicillin sensitive strains, the difference was statistically significant (P0.05); the drug resistance rate among the sampling sites, detecting the presence of certain differences between the.PVL gene and QAC gene: 2 strains of MRSA (2/19 ,10.53%)鍜，

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