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目的研究重型肝炎患者痰中新洋葱伯克霍尔德菌(Burkholderia cenocepacia,BCE)39种耐药基因。方法应用API鉴定条/PSE5.0药敏条和Phoenix NMIC/ID-109鉴定/药敏板鉴定和细菌药敏试验,应用PCR法检测分离于重型肝炎患者合格痰标本1株多药耐药BCE临床分离株16S rRNA、29种β-内酰胺酶相关基因(bla)、6种氨基糖苷类修饰酶基因(AMEs)、消毒剂/磺胺耐药基因(qacE△1-sul1)、3种整合子基因(intⅠ1、2、3)等39种耐药基因,经测序和同源性分析证实并分析其分布情况。结果该菌经16S rRNA测序和同源性分析证实为BCE;药敏试验,对头孢他啶、头孢吡肟、环丙沙星、左氧氟沙星和复方新诺明均敏感,对亚胺培南、美罗培南等均耐药;经测序和同源性分析证实6种耐药基因阳性〔1种bla基因(blaTEM-116)、4种AMEs基因〔aac(6′)-Ⅰb、aac(3)-Ⅰ、ant(2″)-Ⅰ、ant(3″)-Ⅰ〕、和Ⅰ类整合子基因(intI1)〕,blaTEM-116、aac(6′)-Ⅰb、aac(3)-Ⅰ、ant(2″)-Ⅰ、ant(3″)-Ⅰ5种耐药基因GenBank注册号分别为FJ887956、FJ609982、FJ609983、FJ887958、FJ644662;其他28种bla基因、2种AMEs基因(aac(6′)-Ⅱ、aac(3)-Ⅱ)、qacE△1-sul1和2种整合子基因(intⅠ2、intⅠ3)均为阴性。结论该株为多药耐药,其耐药机制为多重机制,主要与6种耐药基因〔blaTEM-116、aac(6′)-Ⅰb、aac(3)-Ⅰ、ant(2″)-Ⅰ、ant(3″)-Ⅰ和intⅠ1〕有关。
Objective To study 39 drug resistance genes of Burkholderia cenocepacia (BCE) in the sputum of patients with severe hepatitis. Methods API identification bar / PSE 5.0 drug sensitive strip and Phoenix NMIC / ID-109 identification / drug susceptibility plate identification and bacterial susceptibility testing were applied. PCR was used to detect the qualified sputum samples from patients with severe hepatitis B multi-drug resistant BCE 16S rRNA, 29 kinds of β-lactamase-related genes (bla), 6 kinds of aminoglycoside-modifying enzyme genes (AMEs), disinfectant / sulfa resistance gene (qacE △ 1-sul1), 3 kinds of integron Genes (intⅠ1,2,3) 39 kinds of resistance genes, confirmed by sequencing and homology analysis and analysis of its distribution. Results The strain was identified as BCE by 16S rRNA sequencing and homology analysis. The susceptibility test was sensitive to ceftazidime, cefepime, ciprofloxacin, levofloxacin and cotrimoxazole, and to imipenem, meropenem, etc. Were all resistant. Six kinds of resistance genes were positive (blaTEM-116), four kinds of AMEs (aac (6 ’) - Ib, aac (2 “) - Ⅰ, ant (3”) - Ⅰ] and class Ⅰ integron gene (intI1)], blaTEM-116, aac (6 ’) - ), Ⅰ, ant (3 “) - Ⅰ, and the GenBank accession numbers of the five resistance genes were FJ887956, FJ609982, FJ609983, FJ887958 and FJ644662 respectively. The other 28 bla genes, two AMEs genes (aac (3) -Ⅱ), qacE △ 1-sul1 and two integron genes (intⅠ2, intⅠ3) were negative. Conclusions The strain is multidrug resistance and its mechanism of multidrug resistance is multifactorial. It is mainly associated with six kinds of resistance genes (blaTEM-116, aac (6 ’) - Ib, aac Ⅰ, ant (3 ”) - Ⅰ and intⅠ1〕.