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用于快速检测新生儿血液中B族链球菌的实时荧光定量PCR检测方法的评估

Evaluation of a real-time fluorescent PCR assay for rapid detection of Group B Streptococci in neonatal blood.

作者信息

Golden Stephen M, Stamilio David M, Faux Brian M, dela Cruz Wilfred P, Shoemaker Craig T, Blackmon Camille L, Stassen Sarah D, Clark Velvet M, Smith James W, Johnson Oswald L

机构信息

Pediatric Flight, David Grant USAF Medical Center, Travis Air Force Base, CA 94535, USA.

出版信息

Diagn Microbiol Infect Dis. 2004 Sep;50(1):7-13. doi: 10.1016/j.diagmicrobio.2004.04.021.

Abstract

Streptococcus agalactiae (Group B Streptococcus: GBS) is the major causative agent of neonatal sepsis. Neonates at risk for GBS infections are empirically administered broad-spectrum antibiotics for at least 48 h pending blood culture results. A rapid assay to expedite detection of GBS would facilitate initiation of specific antibiotic therapy. Conversely, expeditious proof of absence of infection will avoid unnecessary antibiotic use. Using the LightCycler, we evaluated a hybridization probe polymerase chain reaction (PCR) assay to detect GBS-specific cfb gene target DNA sequence in blood specimens. Both sensitivity and specificity of the real-time PCR assay was 100%. The assay demonstrated 100% specificity when tested against 26 non-GBS bacteria. This method is capable of detecting as few as approximately 100 copies or 10 pg of GBS genomic DNA. This real-time PCR method is rapid, sensitive, and specific for the detection of GBS in neonatal blood samples and holds great promise in its utility in the diagnostic laboratory.

摘要

无乳链球菌(B族链球菌:GBS)是新生儿败血症的主要病原体。对于有感染GBS风险的新生儿,在等待血培养结果期间,经验性地给予广谱抗生素治疗至少48小时。一种快速检测GBS的方法将有助于启动特异性抗生素治疗。相反,快速证明没有感染将避免不必要的抗生素使用。我们使用LightCycler评估了一种杂交探针聚合酶链反应(PCR)检测方法,以检测血液标本中GBS特异性cfb基因靶DNA序列。实时PCR检测方法的敏感性和特异性均为100%。在针对26种非GBS细菌进行测试时,该检测方法显示出100%的特异性。该方法能够检测低至约100个拷贝或10 pg的GBS基因组DNA。这种实时PCR方法对于检测新生儿血液样本中的GBS快速、灵敏且特异,在诊断实验室的应用中具有很大前景。

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