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Molecular characterisation of quinolone resistance in Campylobacter jejuni [German]

机译:空肠弯曲菌对喹诺酮耐药的分子特征[德语]

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Increasing occurrence of resistance to fluoroquinolones among Campylobacter from livestock and man has been reported in the last decade. Fluoroquinolone resistance in C. jejuni appears to be due to point mutations in the genes encoding subunits of the DNA gyrase (gyrA) and the topoisomerase IV (parC). The gyrA Thr-86 mutation (ACA --> ATA) is a frequent cause of resistance in C. jejuni isolates and various methods have been reported for the detection of single nucleotide change in Thr-86 of gyrA including, besides direct sequencing, single-strand-conformation polymorphism (SSCP) analysis, mismatch amplification mutation assay (MAMA)-PCR, fluorogenic PCR assay and microarrays (microelectronic chip). PCR-SSCP analysis is a rapid and convenient technique for detection of mutations and allelic variants. Using SSCP-PCR analysis several SSCP patterns with unique nucleic acid sequence combination can be identified. Several silent mutations in different codons from the QRDR (quinolone resistance determining region) of gyrA can also be found. As a screening method SSCP-PCR is suitable for the molecular characterisation of quinolone resistance in C. jejuni, especially because of the low cost and time consumption and easy performance compared to other methods.
机译:在过去的十年中,已经报道了来自家畜和人的弯曲杆菌中对氟喹诺酮类药物的耐药性增加。空肠弯曲杆菌中的氟喹诺酮耐药性似乎是由于编码DNA旋转酶(gyrA)和拓扑异构酶IV(parC)的亚基的基因中的点突变引起的。 gyrA Thr-86突变(ACA-> ATA)是空肠弯曲菌分离株耐药的常见原因,并且已报道了多种检测gyrA Thr-86中单核苷酸变化的方法,除了直接测序外,还包括链构象多态性(SSCP)分析,错配扩增突变测定(MAMA)-PCR,荧光PCR测定和微阵列(微电子芯片)。 PCR-SSCP分析是一种检测突变和等位基因变异的快速便捷的技术。使用SSCP-PCR分析,可以鉴定出具有独特核酸序列组合的几种SSCP模式。还可以从gyrA的QRDR(喹诺酮耐药性决定区域)的不同密码子中找到几个沉默突变。作为一种筛选方法,SSCP-PCR适合空肠弯曲菌对喹诺酮耐药性的分子表征,尤其是因为与其他方法相比,其成本低,耗时少,性能容易的特点。

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