泛耐药鲍曼不动杆菌整合子Ⅰ和ISCR1结构及基因分型研究

目的研究从临床分离的鲍曼不动杆菌的整合子Ⅰ和ISCR1的分布及结构情况,并对其进行基因分型。方法分离自临床的57株鲍曼不动杆菌,PCR检测整合酶Ⅰ、整合子Ⅰ、ISCR1以及ISCR1可变区,PCR产物进行限制性片段长度多态性（RFLP）分型并进行测序分析可变区携带的耐药基因盒,ERIC-PCR进行基因分型。结果 49株整合酶I阳性,其中47株整合子I扩增阳性,RFLP分为2型,测序结果为aacA4-catB8-aadA1和drf17-aadA5。3株ISCR1以及ISCR1可变区扩增均阳性,可变区经RFLP分为1型,测序结果为orf513-qnrA1-ampR-qacEdeltal,ISCR1阳性菌整合子I均阳性,经ERIC-PCR检测将57株鲍曼不动杆菌分为27个基因型。结论Ⅰ类整合子广泛存在于鲍曼不动杆菌中,ISCRI携带率较低,氨基糖苷类、甲氧苄啶类和β-内酰胺类耐药基因盒较常见,ERIC-PCR可用于临床鲍曼不动杆菌的分子流行病学研究。     

产ESBLs、AmpC、KPC酶的葡萄糖不发酵细菌的耐药性分析及临床分布

目的了解本地区葡萄糖不发酵细菌中泛耐药菌（代表株铜绿假单胞菌、鲍曼不动杆菌、嗜麦芽窄食单胞菌）在临床标本中的分离与耐药情况,为临床合理使用抗生素提供指导。方法对临床分离的180株葡萄糖不发酵细菌进行分类鉴定和药敏试验,并用纸片扩散法筛选出产超广谱β-内酰胺酶（ESBLs）、头孢菌素酶（AmpC酶）及碳青霉烯类水解酶（KPC酶）的耐药菌株。结果在葡萄糖不发酵细菌的构成比中,其中产超广谱β-内酰胺酶（ESBLs）56株（31.1%）,产AmpC酶22株（12.2%）,KPC酶2株（1.1%）;同时产ESBLs和AmpC酶16株（8.9%）,同时产ESBLs、AmpC、KPC酶1株（0.6%）。3种主要葡萄糖不发酵细菌对头孢他啶、头孢三嗪、头孢噻肟、哌拉西林的耐药率最高,其中铜绿假单胞菌对阿米卡星、左氧氟沙星、环丙沙星耐药率较低,鲍曼不动杆菌对亚胺培南、左氧氟沙星耐药率较低,嗜麦芽窄食单胞菌对复方新诺明、环丙沙星耐药率较低。结论葡萄糖不发酵细菌中泛耐药菌特别是铜绿假单胞菌、鲍曼不动杆菌、嗜麦芽窄食单胞菌均有较高的多重耐药性,临床上应重视葡萄糖不发酵细菌引起的感染,根据药敏试验结果合理使用抗生素。     

老年人亚胺培南耐药铜绿假单胞菌耐药性研究

目的 明确我院老年病人临床分离铜绿假单胞菌的耐药性、同源性及耐碳青霉烯菌株的基因型。方法 收集我院2006年5月-2009年5月自临床老年病人分离的262株铜绿假单胞菌,纸片扩散法测定其对16种抗菌药物的耐药性;琼脂稀释法和E test法测定耐碳青霉烯菌株对14种抗菌药物的MIC值,PCR扩增及克隆测序分析金属酶基因型。脉冲场凝胶电泳(PFGE)分析携带金属酶基因型菌株的同源性。结果 262株铜绿假单胞菌中筛选到104株耐碳青霉烯。104株耐碳青霉烯铜绿假单胞菌对氨苄西林/舒巴坦、头孢哌酮/舒巴坦两个含舒巴坦制剂药物耐药率分别为78.9％和35.9％,对多黏菌素E耐药率最低为6.0％,对米诺环素耐药率58.3％,其余抗菌药物耐药率均大于70.0％;104株亚胺培南耐药铜绿假单胞菌中12株携带金属酶基因,10株检测到有携带VIM-2基因的1类整合子。PFGE分型中12株菌株属于5个克隆株。结论 在我院流行的亚胺培南耐药铜绿假单胞菌中,金属酶基因不是最主要的基因型,金属β-内酰胺酶均为VIM-2型金属酶,耐药基因盒分布于不同的1类整合子中,整合子播散是最主要的流行方式。     

2014~2016年嗜麦芽窄食单胞菌感染临床分布及耐药性分析

目的 分析我院2014年~2016年所分离嗜麦芽窄食单胞菌在临床标本中的分布和对常用抗菌药物的耐药性。方法 用西门子MicroScan WalkAway96全自动微生物鉴定/药敏测试系统进行细菌鉴定和药物敏感试验,依照CLSI制定的最新判断标准判断药敏试验结果。结果 3年间共检出109株嗜麦芽窄食单胞菌,主要来源于呼吸道痰标本,占83.49%,ICU病区检出最高,占36.70%;其次为急诊科、血液科和神经外科。嗜麦芽窄食单胞菌对复方新诺明和左氧氟沙星敏感性较高,分别为88.99%和79.82%;耐药率较高的是头孢他啶,为74.31%。结论 我院2014年~2016年嗜麦芽窄食单胞菌主要集中在ICU,其次为血液科和神经外科,以呼吸道感染居多。药敏结果显示对复方新诺明和左氧氟沙星敏感性好,替卡西林/棒酸和头孢他啶耐药率较高。     

嗜麦芽窄食单胞菌临床株的多重耐药外排泵的研究

目的　研究嗜麦芽窄食单胞菌临床株外排泵SmeDEF的表达与耐药的关系及其表达调控。方法　琼脂稀释法检测嗜麦芽窄食单胞菌对抗生素敏感性并检测泵抑制剂的作用 ,提取临床菌的RNA进行smeD的RT PCR扩增。提取DNA进行smeT片段的PCR扩增 ,扩增产物进行序列分析。结果　随机选取的 6株嗜麦芽窄食单胞菌均有扩增产物。SmeT的N端氨基酸序列相当保守 ,smeD smeT间区测序发现耐药且泵抑制阳性株基因序列与敏感株明显不同。推测与耐药有关的突变出现在smeT的 82～ 16 5区间。结论　嗜麦芽窄食单胞菌临床株SmeDEF外排泵的表达强弱与其耐药性有关。smeDEF基因的表达可能与调控基因间区的变化有关。     

848株革兰阴性杆菌的临床分布及耐药性分析

目的 了解近年临床分离的革兰阴性杆菌分布及耐药性特征.方法 收集2007年11月～2009年9月临床各类标本中分离的革兰阴性杆菌848株,采用VITEK2全自动细菌鉴定仪进行细菌鉴定及药物敏感实验,用SPSS软件完成数据分析.结果 2007年11月～2009年9月共收集848株革兰阴性杆菌,以大肠埃希菌、肺炎克雷伯菌、铜绿假单胞菌、不动杆菌、阴沟肠杆菌最多见,哌拉西林、环丙沙星、左氧氟沙星、庆大霉素、复方新诺明对大肠埃希菌的MIC90分别为256,8,16,32,640ug/ml.头孢曲松、头孢泊肟、环丙沙星对铜绿假单胞菌的MIC90分别为128,16,8 ug/ml,有较高的耐药性,嗜麦芽窄食单胞菌对所监测的抗生素广谱耐药.结论 临床细菌耐药性日趋严重,有效的进行耐药监测,统计和分析,对强调临床合理应用抗生素有重要的指导意义.     

食品与环境中豚鼠气单胞菌系统发育分析以及毒力和耐药性研究

目的食品与环境中49株豚鼠气单胞菌菌种鉴定、毒力与耐药性检测。方法VITEK、API 20NE生化鉴定,gyrB、rpoD系统发育分析;PCR检测act、alt、ast、lip、ahp、aerA、hlyA、ompA1、fla基因;AST-GN16药敏试验。结果生化鉴定为豚鼠/嗜水气单胞菌株54株,经系统发育分析豚鼠气单胞菌49株,嗜水气单胞菌4株,中国台湾省气单胞菌1株。毒力基因alt、lip、ompA1、fla、act、aerA、hlyA检出率依次为100.00%、100.00%、79.59%、14.29%、2.04%、2.04%、2.04%,未检出ast、ahp;存在4种毒力基因组合,优势组合为alt/lip/ompA1(32/49)。豚鼠气单胞菌氨苄西林、头孢唑林、头孢西丁、阿莫西林/克拉维酸、头孢曲松、复方新诺明、安曲南耐药率依次为79.59%、14.29%、10.20%、6.12%、4.08%、4.08%、2.04%,哌拉西林/他唑巴坦、亚胺培南、阿米卡星、庆大霉素、左氧氟沙星、替加环素、呋喃妥英均敏感;2株多重耐药菌株。结论gyrB、rpoD可有效鉴定豚鼠/嗜水气单胞菌,rpoD可区分近亲缘关系豚鼠与中国台湾省气单胞菌;所有豚鼠气单胞菌均携带2种以上毒力基因,1株环境源菌携带7种毒力基因;青霉素类普遍耐药,产生β-内酰胺酶是最主要耐药机制;未发现碳青霉烯类、氨基糖苷类、氟喹诺酮类、四环素类、呋喃类耐药菌株;食品和生活饮用水存在多重耐药菌。     

396株嗜麦芽窄食单胞菌的耐药特征研究

目的研究嗜麦芽窄食单胞菌的耐药特征。方法采用美国临床实验室标准化委员会（ＮＣＣＬＳ）推荐的纸片扩散法,测定了５年中初次分离的３９６株嗜麦芽窄食单胞菌对ＴＭＰ－ＳＭＺ,多西环素,替卡西林－克拉维酸等２０种抗生素的耐药特征,并对１０例嗜麦芽窄食单胞菌菌血症的治疗进行了分析。结果嗜麦芽窄食单胞菌对ＴＭＰ－ＳＭＺ,替卡西林－克拉维酸,多西环素,环丙沙星和头孢他啶的敏感率最高,分别为８１．０％,９１．０％,９６．０％,７０．７％和５３．８％,而对氨苄西林,氨苄西林－舒巴坦,阿莫西林－克拉维酸,头孢唑林,头孢克罗,头孢美唑,头孢呋辛,头孢噻肟,头孢曲松,氨曲南,亚胺培南高水平耐药,敏感率为（０．９～７．５）％。从替卡西林－克拉维酸,多西环素,ＴＭＰ－ＳＭＺ,环丙沙星和头孢他啶对嗜麦芽窄食单胞菌抑菌环直径分布累积百分率图看出：替卡西林－克拉维酸,多西环素,ＴＭＰ－ＳＭＺ和环丙沙星为活性最好的抗生素。结论替卡西林－克拉维酸,ＴＭＰ－ＳＭＺ和多西环素对所研究的嗜麦芽窄食单胞菌有较高的活性,系嗜麦芽窄食单胞菌菌血症严重感染治疗最有效的抗生素     

一株铜绿假单胞菌中检出两种新的整合子

目的研究铜绿假单胞菌多重耐药的临床分离株RJ217中发现的两个新整合子的结构，并分析其在多重耐药性中的作用。方法用改良三相试验分析RJ217产β-内酰胺酶的情况，用常规和长片段PCR法扩增耐药基因和整合子，并对PCR产物进行序列分析。结果发现铜绿假单胞菌RJ217携带两个新的整合子，其中一个携带veb-I型超广谱β-内酰胺酶(ESBL)基因，这两个整合子的结构分别为IS10-like-veb-I-aadB-oxa10／aadA1和aadB-oxa10／aadA1。结论整合子介导的耐药基因在RJ217的多重耐药性中发挥了重要的作用。     

临床分离133株肺炎克雷伯菌耐药机制和基因分型研究

目的研究临床分离肺炎克雷伯菌的耐药机制及基因多态性。方法用WHONET5．4分析菌株药敏情况。PCR检测菌株I类整合酶、ISCR和ESBLs基因;ERIC—PCR检测基因型,SPSS分析其多态性。结果除亚胺培南和美洛培南,产ESBLs菌的耐药率明显高于不产ESBLs菌（P〈0．005）。对I类整合酶、ISCR、blaTEM、blaSHV、blaCTX-M,88株产ESBLs株的阳性率分别为59．1％、43．2％、69．3％、4．5％、45．5％;45株不产ESBLs株的阳性率分别为40．0％、15．6％、22．2％、4．4％、6．7％。根据指纹图谱把133株肺炎克雷伯菌分为126种,经SPSS系统聚类分析,归为20个大类。结论南方医院产ESBLs肺炎克雷伯菌主要是CTX-M基因型。整合子和ISCR可以共存;ERIC—PCR是一种效果较好的基因分型方法,该院肺炎克雷伯菌呈散发存在。     

60株鲍曼不动杆菌耐药基因携带情况分析

目的研究临床分离的60株鲍曼不动杆菌耐药谱及Ⅰ型整合子和β-内酰胺酶等基因携带情况。方法用微量肉汤稀释法测定16种临床常用抗菌药物的最小抑菌浓度;PCR检测β-内酰胺酶、Ⅰ型整合子和外排泵基因,对阳性基因进行序列分析。结果60株菌中,多重耐药株53株,占88.3%;6株携带OXA-23基因,均对包括碳青霉烯在内的5类以上抗菌药耐药,并具有高耐药特性;38株携带PER-1基因,对头孢菌素类耐药率显著高于PER-1基因阴性菌株(P<0.01);45株检出Ⅰ型整合子结构基因,多重耐药率明显高于Ⅰ型整合子阴性菌株(P<0.01);Ⅰ型整合子和PER-1基因同时阳性25株,与7株两者同为阴性菌株相比,多重耐药率增高(P<0.01),但耐药程度无显著差别。结论Ⅰ型整合子基因及β-内酰胺酶类基因的作用是导致鲍曼不动杆菌多重耐药的重要原因;OXA-23基因阳性菌株多为泛耐药和高耐药株,有必要采取有效措施控制其传播。     

Molecular characterization of multidrug-resistant Salmonella enterica subsp. enterica serovar Typhimurium isolates from swine

As part of a longitudinal study of antimicrobial resistance among salmonellae isolated from swine, we studied 484 Salmonella enterica subsp. enterica serovar Typhimurium (including serovar Typhimurium var. Copenhagen) isolates. We found two common pentaresistant phenotypes. The first was resistance to ampicillin, chloramphenicol, streptomycin, sulfamethoxazole, and tetracycline (the AmCmStSuTe phenotype; 36.2% of all isolates), mainly of the definitive type 104 (DT104) phage type (180 of 187 isolates). The second was resistance to ampicillin, kanamycin, streptomycin, sulfamethoxazole, and tetracycline (the AmKmStSuTe phenotype; 44.6% of all isolates), most commonly of the DT193 phage type (77 of 165 isolates), which represents an unusual resistance pattern for DT193 isolates. We analyzed 64 representative isolates by amplified fragment length polymorphism (AFLP) analysis, which revealed DNA fingerprint similarities that correlated with both resistance patterns and phage types. To investigate the genetic basis for resistance among DT193 isolates, we characterized three AmKmStSuTe pentaresistant strains and one hexaresistant strain, which also expressed resistance to gentamicin (Gm phenotype), all of which had similar DNA fingerprints and all of which were collected during the same sampling. We found that the genes encoding the pentaresistance pattern were different from those from isolates of the DT104 phage type. We also found that all strains encoded all of their resistance genes on plasmids, unlike the chromosomally encoded genes of DT104 isolates, which could be transferred to Escherichia coli via conjugation, but that the plasmid compositions varied among the isolates. Two strains (strains UT08 and UT12) had a single, identical plasmid carrying bla(TEM) (which encodes ampicillin resistance), aphA1-Iab (which encodes kanamycin resistance), strA and strB (which encode streptomycin resistance), class B tetA (which encodes tetracycline resistance), and an unidentified sulfamethoxazole resistance allele. The third pentaresistant strain (strain UT20) was capable of transferring by conjugation two distinct resistance patterns, AmKmStSuTe and KmStSuTe, but the genes were carried on plasmids with slightly different restriction patterns (differing by a single band of 15 kb). The hexaresistant strain (strain UT30) had the same plasmid as strains UT08 and UT12, but it also carried a second plasmid that conferred the AmKmStSuGm phenotype. The second plasmid harbored the gentamicin resistance methylase (grm), which has not previously been reported in food-borne pathogenic bacteria. It also carried the sul1 gene for sulfamethoxazole resistance and a 1-kb class I integron bearing aadA for streptomycin resistance. We also characterized isolates of the DT104 phage type. We found a number of isolates that expressed resistance only to streptomycin and sulfamethoxazole (the StSu phenotype; 8.3% of serovar Typhimurium var. Copenhagen strains) but that had AFLP DNA fingerprints similar or identical to those of strains with genes encoding the typical AmCmStSuTe pentaresistance phenotype of DT104. These atypical StSu DT104 isolates were predominantly cultured from environmental samples and were found to carry only one class I integron of 1.0 kb, in contrast to the typical two integrons (InC and InD) of 1.0 and 1.2 kb, respectively, of the pentaresistant DT104 isolates. Our findings show the widespread existence of multidrug-resistant Salmonella strains and the diversity of multidrug resistance among epidemiologically related strains. The presence of resistance genes on conjugative plasmids and duplicate genes on multiple plasmids could have implications for the spread of resistance factors and for the stability of multidrug resistance among Salmonella serovar Typhimurium isolates.     

Detection and typing of integrons in epidemic strains of Acinetobacter baumannii found in the United Kingdom

Integrons were sought in Acinetobacter isolates from hospitals in the United Kingdom by integrase gene PCR. Isolates were compared by pulsed-field gel electrophoresis, and most belonged to a small number of outbreak strains or clones of A. baumannii, which are highly successful in the United Kingdom. Class 1 integrons were found in all of the outbreak isolates but in none of the sporadic isolates. No class 2 integrons were found. Three integrons were identified among the main outbreak strains and clones. While a particular integron was usually associated with a strain or clone, some members carried a different integron. Some integrons were associated with more than one strain. The cassette arrays of two of the integrons were very similar, both containing gene aacC1, which confers resistance to gentamicin, two open reading frames coding for unknown products (orfX, orfX'), and gene aadA1a, which confers resistance to spectinomycin and streptomycin. The larger of these integrons had two copies of the first (orfX) of the gene cassettes coding for unknown products. The third integron, with a cassette array containing gene aacA4, which codes for amikacin, netilmicin, and tobramycin resistance; a chloramphenicol acetyltransferase, catB8; and gene aadA1, conferring resistance to spectinomycin and streptomycin, was associated with an OXA-23 carbapenemase-producing clone, which has spread rapidly in hospitals in the United Kingdom during 2003 and 2004. These integron cassette arrays have been found in other outbreak strains of A. baumannii from other countries. We conclude that integrons are useful markers for epidemic strains of A. baumannii and that integron typing provides valuable information for epidemiological studies.     

Epidemiological typing of Stenotrophomonas (Xanthomonas) maltophilia by PCR.

We used two PCR methods for epidemiological typing of Stenotrophomonas (Xanthomonas) maltophilia with either arbitrary primers (random amplified polymorphic DNA) or enterobacterial repetitive intergenic consensus sequences as primers (ERIC-PCR). The analysis was performed with 38 isolates of S. maltophilia, comprising 9 nosocomial isolates from a burn unit, 20 other clinical isolates epidemiologically unrelated, and 9 isolates from one cystic fibrosis patient. Both methods indicated that all of the nosocomial episodes were independent. In contrast, the nine isolates from the cystic fibrosis patient were assigned to very closely related profiles, especially by ERIC-PCR. We conclude that random amplified polymorphic DNA and ERIC-PCR have comparable reproducible and discriminatory powers for epidemiological typing of S. maltophilia, but ERIC-PCR profiles can be more easily evaluated.     

沙门菌中第一类整合子的鉴定及特性分析

目的　基于整合子在细菌耐药机制中的重要作用 ,对来自正常人体内耐药沙门菌的整合子分布及其结构特征进行分析。方法　应用PCR方法 ,设计第一类整合酶基因intI1和耐药基因盒的特异性引物 ,用PCR方法检测整合子阳性菌株并对其整合的耐药基因进行测序和序列分析。结果　发现 1株S .hadar和 3株S .tshiongwe为第一类整合酶阳性菌株。耐药基因盒进行扩增的结果 ,从 2株中分别得到 10 0 9bp的扩增产物。 1株得到 16 6 4bp的扩增产物。 1株菌得到 10 0 9bp和 16 6 4bp的扩增产物。序列分析结果表明 ,10 0 9bp的扩增产物为携带aadA2 ,对氨基糖苷类抗生素药物壮观霉素、链霉素产生耐药的基因盒 ;16 6 4bp为携带aadA5和dfr17,对氨基糖苷类抗生素药物壮观霉素、链霉素和磺胺类药物甲氧氨苄嘧啶产生耐药的基因盒。结论　首次揭示了健康人携带由整合子介导的耐药菌这一现象 ,提示我们要从基因水平上监测细菌的耐药情况     

Occurrence of class 1 integrons in uropathogenic fluoroquinolone‐resistant clinical Escherichia coli isolates from Jamaica

Quinolone resistance is generally caused by chromosomal mutations, but has been more recently found associated with the plasmid‐mediated qnr genes. The objective of this study was to screen and analyse polymorphisms of integrons in clinical isolates of Escherichia coli in Jamaica. Previous studies in Jamaica identified fluoroquinolone resistance in predominantly uropathogenic E. coli clinical isolates: 45% harbouring qnrA, qnrB and/or qnrS, and 17% were (Extended‐spectrum beta‐lactamase) ESBL‐producers. These isolates were analysed for the presence and variation of class 1 and 2 integrase genes, 5′‐ and 3′‐ conserved segments and the Orf513 recombinase gene by primer‐specific polymerase chain reaction (PCR) and restriction fragment‐length polymorphism (RFLP). Results indicated integron‐encoded integrases in 93% of isolates primarily harbouring class 1 integrase genes; four of 58 isolates carried both classes. The Orf513 and 5′‐ and 3′‐conserved segment (CS) regions were identified in 83% and 55% of the isolates respectively. RFLP evaluation of the 5′‐ and 3′‐CS regions in int1‐positive strains yielded two main types. The reduced diversity, but wide dispersion of class 1 integrons harbouring qnr genes may give rise to the conservation of the mobile genetic elements in which they are carried.     

Characterization of antimicrobial resistance and class 1 integrons in Enterobacteriaceae isolated from Mediterranean herring gulls (Larus cachinnans)

Mediterranean herring gulls (Larus cachinnans) were investigated as a possible reservoir of antibiotic resistant bacteria and of cassette-borne resistance genes located in class 1 integrons. Two hundred and fourteen isolates of the family Enterobacteriaceae were collected from cloacal swabs of 92 chicks captured in a natural reserve in the North East of Italy. They showed high percentages of resistance to ampicillin and streptomycin. High percentages of resistance to trimethoprim/sulfamethoxazole were found in Proteus and Citrobacter and to chloramphenicol in Proteus. Twenty-two (10%) isolates carried the intI1 gene. Molecular characterization of the integron variable regions showed a great diversity, with the presence of 11 different cassette arrays and of one integron without integrated cassettes. The dfrA1-aadA1a and aadB-aadA2 cassette arrays were the most frequently detected. Also the estX cassette, alone or in combination with other cassettes, was detected in many isolates. From this study it is concluded that the enteric flora of Mediterranean herring gulls may act as a reservoir of resistant bacteria and of resistance genes. Due to their feeding habits and their ability to fly over long distances, these free-living birds may facilitate the circulation of resistant strains between waste-handling facilities, crops, waters, and urban areas.     

Local dissemination of multidrug-resistant Acinetobacter baumannii clones in a Thai hospital

A total of 83 Acinetobacter baumannii isolates from patients attending a tertiary care university hospital in Thailand were investigated for their clonal relatedness, antimicrobial susceptibility profiles, and integron carriage. Susceptibility profiles showed that 56 (67%) of these isolates exhibited multiple drug resistance (MDR). Pulsed-field gel electrophoresis (PFGE) showed that 73% of these resistant isolates were clustered into three predominant PFGE types: 6, 7, and 36. This suggested that the high number of isolates exhibiting MDR phenotypes observed in the hospital is, to some extent, due to the spread of these three resistant clones. Class 1 integrase genes were detected in all MDR isolates belonging to PFGE type 6, most MDR isolates belonging to PFGE type 7 and none of the isolates belonging to PFGE type 36. Five different class 1 gene cassette arrays, dfrA1-orfC, bla(IMP-14)-aac6', aacA4- catB8-aadA1, aacC1-orfX-orfX'-aadA1a, and aacC1-orfX-orfX-orfX'-aadA1a, were identified, of which the bla(IMP-14)-aac6' array has only been found in Thai isolates. Two isolates identified in this study carried a class 2 integrase gene with a 2.2 kb cassette array containing aadA1-sat-dfrA1.