本文選題：沙門氏菌　切入點(diǎn)：耐藥性　出處：《上海交通大學(xué)》2013年碩士論文　論文類型：學(xué)位論文

【摘要】：沙門氏菌是一種重要的食源性致病菌，每年都導(dǎo)致大量的疾病，引起全球范圍內(nèi)的廣泛關(guān)注�？股貙�(duì)致病菌是非常有效的治療手段，但近年來(lái)抗生素的廣泛應(yīng)用使沙門氏菌產(chǎn)生了耐藥性。隨著時(shí)間的推移，耐藥性不斷增強(qiáng)，并且耐藥譜不斷擴(kuò)增，從而增加了對(duì)沙門氏菌感染治療的難度。因此，沙門氏菌的耐藥性受到廣泛的關(guān)注。近年來(lái)發(fā)現(xiàn)，除了基因突變，基因的水平轉(zhuǎn)移也成為病原菌耐藥性傳播重要途徑，例如質(zhì)粒作為獨(dú)立于染色體之外的可移動(dòng)遺傳元件，在耐藥基因的傳播中就起到了不可忽視的作用。因此，分析不同來(lái)源沙門氏菌耐藥性及其可移動(dòng)遺傳元件上耐藥基因攜帶情況，從而揭示可移動(dòng)遺傳元件上耐藥基因與菌株耐藥性的對(duì)應(yīng)關(guān)系具有很重要的意義。 本論文的主要研究?jī)?nèi)容及結(jié)果如下： 1.食源性及醫(yī)源性的沙門氏菌分離株的耐藥性初篩。共選用226株沙門氏菌用試卡法測(cè)試其耐藥性，按2012年美國(guó)實(shí)驗(yàn)室標(biāo)準(zhǔn)委員會(huì)(CLSI)指導(dǎo)原則的標(biāo)準(zhǔn)計(jì)算細(xì)菌對(duì)抗菌藥物的耐藥率(R)，中介率(I)，和敏感率(S)。結(jié)果表明，針對(duì)青霉素類抗生素中，耐氨芐青霉素的菌株達(dá)到15.9%(36/226)，耐舒巴坦/氨芐青霉素達(dá)到12.8%(29/226)；針對(duì)氨基糖苷類抗生素中，耐妥布霉素的菌株達(dá)到6.2%(14/226)、耐慶大霉素5.8%(13/226)；針對(duì)喹諾酮類抗生素中，耐環(huán)丙沙星的菌株達(dá)到4.0%(9/226)，耐左氧氟沙星1.8%(4/226)；針對(duì)頭孢菌素類抗生素中，耐頭孢唑啉4.4%(10/226)，耐頭孢他啶和頭孢曲松分別為2.2%(5/226)，，耐頭孢替坦0.9%(2/226)。針對(duì)碳青霉烯類和除青霉素類、頭孢菌素類以外的β-內(nèi)酰胺酶類，所有分離株的篩查結(jié)果均為敏感。 2.VITEK藥敏實(shí)驗(yàn)結(jié)果中耐藥菌用K-B紙片擴(kuò)散法進(jìn)行復(fù)篩。非敏感表型分離株共59株，亞胺培南、阿米卡星、頭孢吡啶準(zhǔn)符合率(CA)都達(dá)到100%，頭孢他啶標(biāo)準(zhǔn)符合率達(dá)到98.3%，左氧氟沙星為96.6%，頭孢曲松為94.9%，氨曲南準(zhǔn)為93.2%，氨芐青霉素、妥布霉素、環(huán)丙沙星標(biāo)準(zhǔn)符合率達(dá)到91.5%，甲氧胺嘧啶/磺胺甲惡唑?yàn)?6.4%，慶大霉素為84.7%。 3.VITEK試卡法檢測(cè)出來(lái)的非敏感表型的分離株進(jìn)行耐藥基因的篩查。非敏感表型的分離株共84株，分別進(jìn)行了β-內(nèi)酰胺酶類抗生素耐藥基因、喹諾酮類抗生素耐藥基因及氨基糖苷類抗生素耐藥基因的篩查。結(jié)果表明，β-內(nèi)酰胺酶類抗生素非敏感菌株中，β-內(nèi)酰胺酶類耐藥基因攜帶率為59.5%(50/84)，其中CMY基因攜帶率為10.7%(9/84)；OXA基因攜帶率為27.3%(23/84)；TEM基因攜帶率為39.2%(33/84)；PSE基因攜帶率為1.2%(1/84)。對(duì)喹諾酮類抗生素非敏感菌株中，喹諾酮類耐藥基因攜帶率為14.7%(5/34)，其中qnrA基因攜帶率為11.8%(4/34)；qnrS基因攜帶率為2.9%(1/34)。對(duì)喹諾酮類和氨基糖苷類抗生素非敏感菌株中，acc(6’)-Ib-cr基因的攜帶率為17.1%(7/41)。僅CMY基因在食源性和醫(yī)源性分離株中的分布具有統(tǒng)計(jì)學(xué)差異(χ~2=5.480，P0.05)。 4.VITEK試卡法檢測(cè)出來(lái)的非敏感表型的分離株進(jìn)行毒性質(zhì)粒的篩查。對(duì)非敏感表型的分離株共84株進(jìn)行了spv基因的篩查，結(jié)果表明其攜帶率為6.0%(5/84)。這5株沙門氏菌分離株攜帶spv基因全部來(lái)自于醫(yī)源，所以其醫(yī)源性攜帶率為14.7%(5/34)，食源性與醫(yī)源性分離株的攜帶率無(wú)統(tǒng)計(jì)學(xué)差異(χ~2=2.05，P0.05)。 利用本研究獲得的食源性和醫(yī)源性沙門氏菌耐藥表型及耐藥基因型信息，可為食品安全風(fēng)險(xiǎn)評(píng)估提供依據(jù)，同時(shí)為臨床治療用藥選擇提供參考數(shù)據(jù)。
[Abstract]:Salmonella is an important foodborne pathogen, has led to a large number of diseases each year, causing widespread concern worldwide. Antibiotics are very effective treatment of pathogens, but the extensive application of antibiotics in recent years that Salmonella produced resistance. With the passage of time, resistance increased, and drug resistance continuous spectrum amplification, thus increasing the difficulty of treatment in Salmonella infection. Therefore, the drug resistance of Salmonella has been widely concerned. In recent years, in addition to gene mutation, gene horizontal transfer of antibiotic resistance of pathogens has become an important means of communication, such as a plasmid as an independent chromosome outside of mobile genetic elements, not played an essential role in the spread of resistance genes. Therefore, analysis of drug resistance of Salmonella from different sources and mobile genetic elements on resistance gene carrying case, It is of great significance to reveal the relationship between the resistance genes of the mobile genetic elements and the resistance of the strains.
The main contents and results of this paper are as follows:
1. foodborne and iatrogenic Salmonella isolates resistance screening. A total of 226 strains of Salmonella by test method to test the resistance of the United States in 2012, according to the Laboratory Standards Committee (CLSI) the guiding principles of the standard to calculate the antibiotic resistance rate (R), intermediate rate (I), and sensitive rate (S). The results show that for penicillin, ampicillin resistant strains reached 15.9% (36/226), resistance to ampicillin / sulbactam reached 12.8% (29/226); for aminoglycosides, tobramycin resistant strains reached 6.2% (14/226), 5.8% (13/226); gentamicin resistance to quinolones antibiotics, ciprofloxacin resistant strains reached 4% (9/226), levofloxacin resistant (4/226); according to the 1.8% cephalosporin antibiotics, resistant to cefazolin (4.4% 10/226), resistant to ceftazidime and ceftriaxone were 2.2% (5/226), resistance to cefotetan (0.9% 2/226). The screening results of all the isolates were sensitive to the beta lactamases other than cephalosporins for carbapenems and penicillins.
The experimental results of 2.VITEK drug sensitivity in drug resistant bacteria by K-B disk diffusion method for multiple screen. Non sensitive phenotype isolates there were 59 strains of imipenem, Amikacin, ceftazidime pyridine (CA) quasi coincidence rate reached 100%, ceftazidime standard rate of 98.3%, levofloxacin 96.6%, head of ceftriaxone was 94.9%, ammonia aztreonam quasi 93.2%, ampicillin, ciprofloxacin and tobramycin, the standard rate of 91.5%, methoxamine pyrimidine / sulfamethoxazole gentamicin was 86.4%, 84.7%.
3.VITEK test method to detect non sensitive isolates were screened for resistance genes. Non sensitive isolates of 84 strains were beta lactam antibiotic resistance genes, screening of quinolone antibiotic resistance genes and aminoglycoside antibiotic resistance genes. The results showed that beta lactam non sensitive isolates, beta lactamase resistant gene carrying rate was 59.5% (50/84), the CMY gene carrying rate was 10.7% (9/84); OXA gene carrying rate was 27.3% (23/84); TEM gene carrying rate was 39.2% (33/84); PSE gene carrying rate was 1.2% (1/84) of quinolone antibiotics non sensitive isolates, quinolone resistance gene carrying rate was 14.7% (5/34), the qnrA gene carrying rate was 11.8% (4/34); qnrS gene carrying rate was 2.9% (1/34). To quinolones and aminoglycosides (non sensitive strains, ACC The carrying rate of the 6 '-Ib-cr gene was 17.1% (7/41). The distribution of only CMY gene in food borne and iatrogenic isolates was statistically different (x ~2=5.480, P0.05).
4.VITEK test method to detect non sensitive isolates were screened for virulence plasmid. The non sensitive isolates were screened 84 strains of SPV gene, the results showed that the positive rate is 6% (5/84). The 5 strains of Salmonella strains carrying SPV gene from all medical sources, so the iatrogenic carrying rate was 14.7% (5/34), and the medical source of foodborne isolates carrying rate showed no significant difference (~2=2.05, P0.05).
Using the information obtained from this study on drug resistance phenotype and resistance genotype of Salmonella, we can provide a basis for food safety risk assessment, and provide reference data for clinical treatment.

【學(xué)位授予單位】：上海交通大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2013
【分類號(hào)】：R3416

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