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首页> 外文OA文献 >Simultaneous Detection of Nine Antibiotic Resistance-Related Genes in Streptococcus agalactiae Using Multiplex PCR and Reverse Line Blot Hybridization Assay†
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Simultaneous Detection of Nine Antibiotic Resistance-Related Genes in Streptococcus agalactiae Using Multiplex PCR and Reverse Line Blot Hybridization Assay†

机译：使用多重PCR和逆线印迹杂交法同时检测无乳链球菌中九种抗生素抗药性相关基因†

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Streptococcus agalactiae (group B streptococcus [GBS]) is the leading cause of neonatal and maternal sepsis. Penicillin is recommended for intrapartum prophylaxis, but erythromycin or clindamycin is used for penicillin-allergic carriers. Antibiotic resistance (AR) has increased recently and needs to be monitored. We have developed a multiplex PCR-based reverse line blot (mPCR/RLB) hybridization assay to detect, simultaneously, seven genes encoding AR—erm(A/TR), erm(B), mef(A/E), tet(M), tet(O), aphA-3, and aad-6—and two AR-related genes, int-Tn and mreA. We tested 512 GBS isolates from Asia and Australasia and compared mPCR/RLB with antibiotic susceptibility phenotype or single-gene PCR. Phenotypic resistance to tetracycline was identified in 450 (88%) isolates, of which 442 had tet(M) (93%) and/or tet(O) (6%). Of 67 (13%) erythromycin-resistant isolates, 18 were susceptible to clindamycin, i.e., had the M phenotype, encoded by mef(A/E); 39 had constitutive (cMLSB) and 10 inducible clindamycin resistance, and of these, 34 contained erm(B) and 12 erm(A/TR). Of four additional isolates with mef(A/E), three contained erm(B) with cMLSB and one was erythromycin susceptible. Of 61 (12%) clindamycin-resistant isolates, 20 were susceptible to erythromycin and two had intermediate resistance. Based on sequencing, 21 of 22 isolates with mef had mef(E), and 8 of 353 with int-Tn had an atypical sequence. Several AR genes, erm(B), tet(O), aphA-3, aad-6, and mef(A/E), were significantly more common among Asian than Australasian isolates, and there were significant differences in distribution of AR genes between GBS serotypes. Our mPCR/RLB assay is simple, rapid, and suitable for surveillance of antibiotic resistance in GBS.
机译：无乳链球菌（B组链球菌[GBS]）是新生儿和产妇败血症的主要原因。建议使用青霉素预防产时，但将红霉素或克林霉素用于对青霉素过敏的载体。抗生素耐药性（AR）最近有所增加，需要进行监测。我们已经开发了一种基于多重PCR的逆线印迹（mPCR / RLB）杂交测定法，以同时检测编码AR-erm（A / TR），erm（B），mef（A / E），tet（M）的七个基因），tet（O），aphA-3和aad-6以及两个与AR相关的基因int-Tn和mreA。我们测试了来自亚洲和大洋洲的512 GBS分离株，并将mPCR / RLB与抗生素敏感性表型或单基因PCR进行了比较。在450个（88％）分离物中鉴定出对四环素的表型抗性，其中442个具有tet（M）（93％）和/或tet（O）（6％）。在67株（占13％）抗红霉素菌株中，有18株对克林霉素敏感，即具有由mef（A / E）编码的M表型。 39个具有组成型（cMLSB）和10个诱导的克林霉素抗性，其中34个包含erm（B）和12 erm（A / TR）。在另外四个带有mef（A / E）的分离株中，三个带有cMLSB的erm（B）分离株，一个对红霉素敏感。在对克林霉素有抗药性的61株（占12％）中，有20株对红霉素敏感，有2株具有中等耐药性。根据测序结果，具有mef的22个分离株中有21个具有mef（E），具有int-Tn的353个分离株中有8个具有非典型序列。几个AR基因，erm（B），tet（O），aphA-3，aad-6和mef（A / E）在亚洲人中比在澳大利亚分离株中更为常见，并且AR基因的分布存在显着差异在GBS血清型之间。我们的mPCR / RLB检测方法简单，快速，适合监测GBS中的抗生素耐药性。

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