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首页> 外文期刊>Journal of Clinical Microbiology >Development of a Rapid PCR Assay Specific forStaphylococcus saprophyticus and Application to Direct Detection from Urine Samples
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Development of a Rapid PCR Assay Specific forStaphylococcus saprophyticus and Application to Direct Detection from Urine Samples

机译:腐生葡萄球菌特异快速PCR检测方法的开发及其在尿液样品中直接检测的应用

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Staphylococcus saprophyticus is one of the most frequently encountered microorganisms associated with acute urinary tract infections (UTIs) in young, sexually active female outpatients. Conventional identification methods based on biochemical characteristics can efficiently identify S. saprophyticus, but the rapidities of these methods need to be improved. Rapid and direct identification of this bacterium from urine samples would be useful to improve time required for the diagnosis of S. saprophyticus infections in the clinical microbiology laboratory. We have developed a PCR-based assay for the specific detection ofS. saprophyticus. An arbitrarily primed PCR amplification product of 380 bp specific for S. saprophyticus was sequenced and used to design a set of S. saprophyticus-specific PCR amplification primers. The PCR assay was specific for S. saprophyticus when tested with DNA from 49 gram-positive and 31 gram-negative bacterial species. This assay was also able to amplify efficiently DNA from all 60 strains of S. saprophyticus from various origins tested. This assay was adapted for direct detection from urine samples. The sensitivity levels achieved with urine samples was 19 CFU with 30 cycles of amplification and 0.5 CFU with 40 cycles of amplification. This PCR assay for the specific detection of S. saprophyticus is simple and rapid (approximately 90 min, including the time for urine specimen preparation).
机译:葡萄球菌是年轻的,有性活跃的女性门诊患者中最常见的与急性尿路感染(UTI)相关的微生物之一。基于生化特性的常规识别方法可以有效地识别 S。腐生菌,但是这些方法的快速性有待提高。从尿液样本中快速,直接鉴定该细菌将有助于缩短诊断 S所需的时间。在临床微生物学实验室感染腐生菌。我们已经开发了一种基于PCR的检测方法,用于特异性检测 S。腐生菌。对 S具有特异性的380 bp的任意引物PCR扩增产物。对腐生菌进行测序,并用于设计一组 S。腐生菌特异的PCR扩增引物。 PCR分析对 S是特异的。用49克阳性和31克阴性细菌的DNA检测腐生菌。该测定法还能够有效扩增来自所有60株 S的DNA。测试了来自不同来源的腐生菌。该测定适用于直接从尿液样品中检测。尿液样品达到的敏感度水平为19 CFU(30个扩增循环)和0.5 CFU(40个扩增循环)。此PCR检测法用于特异性检测 S。腐生菌简单,快速(约90分钟,包括尿液标本制备时间)。

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