常见革兰阴性杆菌耐药性和消毒剂—磺胺耐药基因的检测及分析
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摘要
目的了解临床常见革兰阴性杆菌分离菌株对常用抗菌药物的耐药现状及其抗消毒剂基因的携带情况,为临床治疗感染性疾病提供科学依据。
方法采用BD Phoenixl00全自动微生物分析仪鉴定细菌；药敏试验采用K-B琼脂纸片扩散法；聚合酶链反应(PCR)检测抗消毒剂基因和磺胺耐药基因(qacEΔ1-sull)。
结果①临床共分离208株革兰阴性杆菌,其中大肠埃希菌49株,肺炎克雷伯菌53株,阴沟肠杆菌10株,铜绿假单胞菌48株,鲍曼不动杆菌48株。主要分布于ICU、呼吸科、肾内科、老年内科和神经科病房,不同细菌在不同科室分布有差别。②临床常用抗生素中,肠杆菌科临床分离菌株耐药率低于20%的抗生素有阿米卡星、亚胺培南、美洛培南、哌拉西林/他唑巴坦和头孢哌酮/舒巴坦；耐药率高于80%的抗生素有氨苄西林。③临床分离的非发酵菌菌株中,铜绿假单胞菌耐药率低于30%的抗生素有阿米卡星、妥布霉素、哌拉西林/他唑巴坦和头孢哌酮/舒巴坦；耐药率高于90%的抗生素有头孢唑林、头孢呋肟、氨苄西林、氨苄西林/舒巴坦和呋喃妥因。鲍曼不动杆菌对抗生素的耐药水平普遍较高,耐药率低于45%的抗菌药物有哌拉西林/他唑巴坦和头孢哌酮/舒巴坦。耐药率高于90%的抗菌药物有头孢唑林、头孢呋肟和呋喃妥因。④208例革兰阴性杆菌分离菌株中,有104株携带qacE△1-sull基因,总阳性率为50%。其中鲍曼不动杆菌中携带qacE△1-sull基因阳性率高达75%,其次是大肠埃希氏菌为69.4%,2种菌株qacE△1-sull基因携带率明显高于其它菌株(P<0.05)。⑤各科室中,ICU检出携带qacEΔ1-sull基因革兰阴性杆菌42株,阳性率高达71.2%,明显高于呼吸科和神经科菌株的携带率(P<0.05)。
结论①ICU病房是常见革兰阴性杆菌分布较高的科室,提示免疫力低下、使用呼吸机、行气管插管等侵入性操作的患者以及长期住院使用抗菌药物治疗的患者应密切注意革兰阴性杆菌等条件致病菌引起的感染。②肠杆菌科中大肠埃希氏菌和肺炎克雷伯菌多重耐药严重,亚胺培南和美洛培南仍是治疗大肠埃希菌和肺炎克雷伯菌感染的首选药物,抗生素复合制剂头孢哌酮/舒巴坦也是治疗大肠埃希氏菌和肺炎克雷伯菌感染的理想药物。③非发酵菌中铜绿假单胞菌和鲍曼不动杆菌对常用抗生素呈高水平耐药,哌拉西林/他唑巴坦和头孢哌酮/舒巴坦对治疗二者感染有较好效果。④革兰阴性杆菌临床分离菌株qacEΔ1-sull基因携带率较高,以鲍曼不动杆菌和大肠埃希菌尤为明显,因此应加强对多重耐药菌株的耐药性监测,重视抗菌药物和医用消毒剂的使用。
Objective:To investigate the drug resistance of common gram-negative bacilli to regular antibiotics and detect the disinfectant-resistant genes, providing a scientific basis for clinical treatment of infectious diseases.
Methods:All isolated strains were identified by BD Phoenix automated microbiology system (BD Phoenix100). Antibiotic sensitivity analysis of the isolated strains were taken by Kirby-Bauer method, and the disinfectant-resistant gene (qacEΔ1-sull) was detected by polymerase chain reaction (PCR).
Results:①208gram-negative bacilli were isolated from clinical specimens, including Escherichia coli (49strains), Klebsiella pnenmoniae(53strains), Enterobacter cloacae(10strains), verdigris Pseudomonca (48strains) and Acinetobacter baumannii(48strains), which were mainly distributed in intensive care unit (ICU), respiratory and neurological department.②Among the commonly used antibiotics in clinical treatments, the drug resistance rates of enterobacteria to Amikacin, Imipenem, Meropenem, Piperacillin/Tazobactam and Cefoperazone/Sulbactam were less than20%, while the rate to Ampicillin was higher than80%.③Among the isolated non-fermentative bacteria, the resistance rates of verdigris Pseudomonca to Amikacin, Tobramycin, Piperacillin/Tazobactam and Cefoperazone/Sulbactam were less than30%. while the resistance rates to Cefazolin, Cefuroxime, Ampicillin. Ampicillin/Sulbactam and Nitrofurantoin were higher than90%. Moreover, drug resistance of Acinetobacter baumannii was strictly high. To Piperacillin/Tazobactam and Cefoperazone/Sulbactam the rates were less than45%, while to Cefazolin, Cefuroxime and Nitrofurantoin, the rates were more than90%.④The104out of208(50%) strains carried qacEΔ1-sull gene detected by PCR. The positive rate of Acinetobacter baumannii carrying qacEA1-sull gene that was the highest (75%). Escherichia coli was69.4%. The positive rate of Acinetobacter baumannii and Escherichia coli carrying qacEAl-sull gene were higher than other bacteria strains(P<0.05).⑤The42strains carrying qacEΔ1-sull gene out of208(71.2%) strains in ICU were identified, which was obviously higher than respiratory and neurological department(P<0.05).
Conclusions:①mong all the departments, the distribution of gram-negative bacilli from ICU is the highest, suggesting that patients in ICU with poor immunity, using ventilator or receiving invasive operations like tracheal intubation and taking antibiotics for a long time should be paid more attention to the infection of gram-negative bacilli.②Escherichia coli and Klebsiella pnenmoniae (belonging to enterobacteria) are severely muti-drug resistant, but Imipenem and Meropenem are still the first choice to treat infections of Escherichia coli and Klebsiella pnenmoniae.③Verdigris Pseudomonca and Acinetobacter baumannii (belonging to non-fermentative bacteria) are highly resistant to common antibiotics, while their infections can be effectively treated by Piperacillin/Tazobactam and Cefoperazone/Sulbactam.④Among the isolated gram-negative bacilli, the positive rate of qacEAl-sull gene is high, especially Verdigris Pseudomonca and Escherichia coli, indicating that the usage of antibiotics and disinfectants should be more strictly controlled.
方法采用BD Phoenixl00全自动微生物分析仪鉴定细菌；药敏试验采用K-B琼脂纸片扩散法；聚合酶链反应(PCR)检测抗消毒剂基因和磺胺耐药基因(qacEΔ1-sull)。
结果①临床共分离208株革兰阴性杆菌,其中大肠埃希菌49株,肺炎克雷伯菌53株,阴沟肠杆菌10株,铜绿假单胞菌48株,鲍曼不动杆菌48株。主要分布于ICU、呼吸科、肾内科、老年内科和神经科病房,不同细菌在不同科室分布有差别。②临床常用抗生素中,肠杆菌科临床分离菌株耐药率低于20%的抗生素有阿米卡星、亚胺培南、美洛培南、哌拉西林/他唑巴坦和头孢哌酮/舒巴坦；耐药率高于80%的抗生素有氨苄西林。③临床分离的非发酵菌菌株中,铜绿假单胞菌耐药率低于30%的抗生素有阿米卡星、妥布霉素、哌拉西林/他唑巴坦和头孢哌酮/舒巴坦；耐药率高于90%的抗生素有头孢唑林、头孢呋肟、氨苄西林、氨苄西林/舒巴坦和呋喃妥因。鲍曼不动杆菌对抗生素的耐药水平普遍较高,耐药率低于45%的抗菌药物有哌拉西林/他唑巴坦和头孢哌酮/舒巴坦。耐药率高于90%的抗菌药物有头孢唑林、头孢呋肟和呋喃妥因。④208例革兰阴性杆菌分离菌株中,有104株携带qacE△1-sull基因,总阳性率为50%。其中鲍曼不动杆菌中携带qacE△1-sull基因阳性率高达75%,其次是大肠埃希氏菌为69.4%,2种菌株qacE△1-sull基因携带率明显高于其它菌株(P<0.05)。⑤各科室中,ICU检出携带qacEΔ1-sull基因革兰阴性杆菌42株,阳性率高达71.2%,明显高于呼吸科和神经科菌株的携带率(P<0.05)。
结论①ICU病房是常见革兰阴性杆菌分布较高的科室,提示免疫力低下、使用呼吸机、行气管插管等侵入性操作的患者以及长期住院使用抗菌药物治疗的患者应密切注意革兰阴性杆菌等条件致病菌引起的感染。②肠杆菌科中大肠埃希氏菌和肺炎克雷伯菌多重耐药严重,亚胺培南和美洛培南仍是治疗大肠埃希菌和肺炎克雷伯菌感染的首选药物,抗生素复合制剂头孢哌酮/舒巴坦也是治疗大肠埃希氏菌和肺炎克雷伯菌感染的理想药物。③非发酵菌中铜绿假单胞菌和鲍曼不动杆菌对常用抗生素呈高水平耐药,哌拉西林/他唑巴坦和头孢哌酮/舒巴坦对治疗二者感染有较好效果。④革兰阴性杆菌临床分离菌株qacEΔ1-sull基因携带率较高,以鲍曼不动杆菌和大肠埃希菌尤为明显,因此应加强对多重耐药菌株的耐药性监测,重视抗菌药物和医用消毒剂的使用。
Objective:To investigate the drug resistance of common gram-negative bacilli to regular antibiotics and detect the disinfectant-resistant genes, providing a scientific basis for clinical treatment of infectious diseases.
Methods:All isolated strains were identified by BD Phoenix automated microbiology system (BD Phoenix100). Antibiotic sensitivity analysis of the isolated strains were taken by Kirby-Bauer method, and the disinfectant-resistant gene (qacEΔ1-sull) was detected by polymerase chain reaction (PCR).
Results:①208gram-negative bacilli were isolated from clinical specimens, including Escherichia coli (49strains), Klebsiella pnenmoniae(53strains), Enterobacter cloacae(10strains), verdigris Pseudomonca (48strains) and Acinetobacter baumannii(48strains), which were mainly distributed in intensive care unit (ICU), respiratory and neurological department.②Among the commonly used antibiotics in clinical treatments, the drug resistance rates of enterobacteria to Amikacin, Imipenem, Meropenem, Piperacillin/Tazobactam and Cefoperazone/Sulbactam were less than20%, while the rate to Ampicillin was higher than80%.③Among the isolated non-fermentative bacteria, the resistance rates of verdigris Pseudomonca to Amikacin, Tobramycin, Piperacillin/Tazobactam and Cefoperazone/Sulbactam were less than30%. while the resistance rates to Cefazolin, Cefuroxime, Ampicillin. Ampicillin/Sulbactam and Nitrofurantoin were higher than90%. Moreover, drug resistance of Acinetobacter baumannii was strictly high. To Piperacillin/Tazobactam and Cefoperazone/Sulbactam the rates were less than45%, while to Cefazolin, Cefuroxime and Nitrofurantoin, the rates were more than90%.④The104out of208(50%) strains carried qacEΔ1-sull gene detected by PCR. The positive rate of Acinetobacter baumannii carrying qacEA1-sull gene that was the highest (75%). Escherichia coli was69.4%. The positive rate of Acinetobacter baumannii and Escherichia coli carrying qacEAl-sull gene were higher than other bacteria strains(P<0.05).⑤The42strains carrying qacEΔ1-sull gene out of208(71.2%) strains in ICU were identified, which was obviously higher than respiratory and neurological department(P<0.05).
Conclusions:①mong all the departments, the distribution of gram-negative bacilli from ICU is the highest, suggesting that patients in ICU with poor immunity, using ventilator or receiving invasive operations like tracheal intubation and taking antibiotics for a long time should be paid more attention to the infection of gram-negative bacilli.②Escherichia coli and Klebsiella pnenmoniae (belonging to enterobacteria) are severely muti-drug resistant, but Imipenem and Meropenem are still the first choice to treat infections of Escherichia coli and Klebsiella pnenmoniae.③Verdigris Pseudomonca and Acinetobacter baumannii (belonging to non-fermentative bacteria) are highly resistant to common antibiotics, while their infections can be effectively treated by Piperacillin/Tazobactam and Cefoperazone/Sulbactam.④Among the isolated gram-negative bacilli, the positive rate of qacEAl-sull gene is high, especially Verdigris Pseudomonca and Escherichia coli, indicating that the usage of antibiotics and disinfectants should be more strictly controlled.
引文
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[11]Mac Gowan AP, Bowker KE. Mechanism of fluoroquinolone resistanceis an important factor in determining the antimicrobial effect of gemifloxacin against Streptoco-ccus pneumoniae in an in vitro pharmaco kinetic model. Antimicrob Agents Chemother, 2003,47 (3):1096.
[12]冯兴军,王建华,单安山.抗菌肽基因工程研究及表达策略.中国生物工程 杂志,2006,26(3)：63-67.
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[16]陆建荣,王惠民,吴萍,黄松平,常秋月,凌勇武,倪晓蓉,5'-非转录区序列改建提高毕赤酵母表达抗菌肽LL-37.第二军医大学学报,2007,28(12)：1329-1334.
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[18]尹娜,李鸿钧,彭梅,谢天宏,张光明,施锐,孙茂盛.抗菌肽Cecropin D在毕赤酵母中的表达、纯化及活性鉴定.中国生物制品学杂志,2008,21(3)：185-189.
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[20]Li XZ Ma D, Livermore DM, Nikaido H. Role of efflux pump(s) in intrinsic resistance of Pseudomonas aeruginosa:active efflux as a contributing factor to beta-lactam resistance. Antimicrob Agents Chemother,1994,38:1742
[21]周黎明,王正荣,王浴生.大肠杆菌主动外排泵与抗生素多重耐药性研究进展.四川生理科学杂志,2003,25(4)：163-164.
[22]张海旺,邓旭明,任晓慧,唐峰,褚秀玲.主动外排系统介导的大肠杆菌多重耐药性的确证.中国兽医学报,2005,25(2)：173-175.
[23]赵廷坤,凌保东,周歧新.铜绿假单胞菌主动外排系统研究进展.四川生理科学杂志,2004,26(3)：126-130.
[24]宋战昀,韩文瑜,雷连成,冯新.枯草杆菌多重耐药基因bmr的克隆及原核表达.中国兽医学报,2005,25(6)：597-599.
[25]汀复.抗菌药物合理应用的儿个问题.中国抗感染化疗杂志,2005,5(1)：1-3.
[26]王丽,杨永弘,陆权,王艺,陈沅,邓力,邓秋莲,张泓,王传清,刘岚,徐樨魏,沈叙庄.儿科常见革兰阴性杆菌耐药性现状分析.中国感染与化疗杂志,2008,8(3)：177-182.
[27]汀复,朱德妹,胡付品,阮斐怡,倪语星,孙景勇,徐英春,张小江,胡云健,艾效曼, 俞云松,杨青,孙自镛,简翠,贾蓓,黄文祥,卓超,苏丹虹,魏莲花,吴玲,张朝霞,季萍,王传清,薛建昌,张泓,李万华.2008年中国CHINET细菌耐药性监测.中国感染与化疗杂志,2009,9(5)：321-329.
[28]侯丽影,郑跃杰,邓继岿,赵瑞珍.深圳市儿童社区获得性肺炎细菌病原学及其耐药性.中国微生态学杂志,2008,20(6)：586-588.
[29]许锦民,苏丹虹.大肠埃希菌和克雷伯菌属产超广谱p-内酰胺酶的检测.中华医学检验杂志,1998,21(6)：373.
[30]汪复.多重耐药铜绿假单胞菌与鲍曼不动杆菌严重感染的防治策略.中国感染与化疗杂志,2007,7(3)：230-232.
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[32]余秉翔,余丹阳,马家兰,刘又宇.高产AmpC酶阴沟肠杆菌感染的临床危险因系分析.中华医院感染学杂志,2003,13(6)：511-513.
[33]Zhou Z, Li L, Yu Y, Ma Y. The status of drug resistance and AmpC gene expression in Enterobacter cloacae. Chinese Med J,2003,116(8):1244-1247.
[34]李湘燕,肖永红Mohnarin 2006-2007年度报告：14岁以下人群细菌耐药监测.中国抗生素杂志,2008,33(10)：579-585.
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