Application of six multiplex PCR's among 200 clinical isolates of Pseudomonas aeruginosa for the detection of 20 drug resistance encoding genes
Application of six multiplex PCR's among 200 clinical isolates of Pseudomonas aeruginosa for the detection of 20 drug resistance encoding genes
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DOI：
10.1016/j.kjms.2017.09.010
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发表时间：
2018-02-01
影响因子：
3.3
通讯作者：
Madhavan, Hajib NarahariRao
中科院分区：
文献类型：
--
作者：
Murugan, Nandagopal;Malathi, Jambulingam;Madhavan, Hajib NarahariRao
铜绿假单胞菌（P. aeruginosa，P. aeruginosa）是一种严重的机会致病菌，其多重耐药（multidrug resistant，MDRPA）机制与β-内酰胺酶、孔蛋白基因抑制和外排泵过表达有关。本研究的重点是标准化和应用多重PCR（mPCR）检测编码bla（Tem）、bla（OXA）、bla（CTX-M-15）、bla（Vim）、bla（Ges）、bla（Veb）、bla（DIM）的β内酰胺酶基因和编码Mex A、B-oprM、Mex C、D-oprJ、Mex X、Y-oprN、oprD、nfxB、MexR的外排泵基因的存在。通过mPCR对总共200株铜绿假单胞菌临床分离株进行基因型检测，并通过表型方法对ESBL和MBL产生进行表征。在200个分离株中，163个nfxB调节基因102例（51%），MexC 96例（48%），MexB基因46.5%，MexD基因43%，OprM基因40.5%，OprJ基因37%，OprD + MexR基因36%，MexX + OprN基因26.5%，MexY基因24.5%。mPCR检测β内酰胺酶基因bla（Tem）145（72.5%）、bla（OXA）67（33.5%）、blaVim 35（17.5%）、blaVeb 25（12.50%）、23（11.50%）、blaGes 21（11.5%）、Ctx-m 14（7%）、AmpC 10（5%）、blaDim-1 5（2.5%）。200例ESBL和MBL表型阳性者分别为38例（19%）和29例（14.5%）。应用该mPCR技术对临床标本进行筛选是一种快速、准确、特异、低成本的可靠工具，其中培养阴性的真细菌PCR阳性病例可用于早期分子检测耐药机制，辅助临床医生选择合适的抗生素治疗疾病。版权所有（C）2017，高雄医学大学.出版社：Elsevier Taiwan LLC
Pseudomonas aeruginosa (P. aeruginosa) is a menacing opportunistic, nosocomial pathogen; become a growing concern as conventional antimicrobial therapy is now futile against it. Multi-drug resistant P. aeruginosa (MDRPA) has distinctive resistance mechanisms such as production of b-lactamases, repression of porin genes and over-expression of efflux pumps. The focus of this study is to standardize and application of multiplex PCR (mPCR) to detect the presence of betalactamase genes encoding bla(Tem), bla(OXA), bla(CTX-M-15), bla(Vim), bla(Ges), bla(Veb), bla(DIM), AmpC and Efflux pump genes encoding Mex A, B-oprM, Mex C, D-oprJ, Mex X, Y-oprN, oprD, nfxB, MexR. A total of 200 clinical isolates of P. aeruginosa were tested for the presence of the above mentioned genes genotypically through mPCR and characterized by phenotypic methods for ESBL and MBL production. Out of 200 isolates, 163 (81.5%) nfxB regulator gene, 102 (51%) MexA, 96 (48%) MexC, 93 (46.5%) MexB, 86 (43%) MexD, 81 (40.5%) OprM, 74 (37%) OprJ, 72 (36%) OprD and MexR, 53 (26.5%) Mex X and OprN, 49 (24.5%) MexY gene. Betalactamase genes 145 (72.5%) bla(Tem), 67 (33.5%) bla(OXA), 35 (17.5%) blaVim, 25(12.50%), 23 (11.50%) blaVeb, 21 (11.5%) blaGes, 14 (7%) Ctx-m and 10 (5%) AmpC and 5 (2.5%) blaDim-1 gene were tested positive by mPCR. Phenotypically 38 (19%) and 29 (14.5%) out of 200 tested positive for ESBL and MBL production. Application of this mPCR on clinical specimens is fast, accurate, specific and low-cost reliable tool for the screening, where culture negative Eubacterial PCR positive cases for an early molecular detection of drug resistance mechanism assisting the clinician to treat the disease with appropriate antibiotic selection. Copyright (C) 2017, Kaohsiung Medical University. Published by Elsevier Taiwan LLC.