评价碱性洗脱/裂解程序在临床常规实践中对阳性血培养的分子诊断价值。
Evaluation of the alkaline wash/lysis procedure for the molecular diagnosis of a positive bacterial blood culture in clinical routine practice.
机构信息
Hsin Sheng College of Medical Care and Management, Taoyuan, Taiwan.
出版信息
J Clin Lab Anal. 2010;24(3):139-44. doi: 10.1002/jcla.20396.
Abstract
Blood culture is commonly used to detect microorganisms in patients with a suspected blood infection. This study evaluated the alkaline wash/lysis procedure to extract DNA of microorganisms in a clinical blood culture. A multiplex polymerase chain reaction (PCR) targeting the 16S rDNA (ribosomal DNA) gene and the fungal ITS (internal transcribed spacer) gene was used as a reliable indicator for the presence of microorganism DNA in the extracts. A total of 535 BacT/ALERT positive blood culture bottles were evaluated. Multiplex PCR showed positive results in 530 DNA extracts, but 5 DNA extracts gave negative results. We conclude that the alkaline wash/lysis procedure in combination with the multiplex PCR is a simple and sensitive method, which can be used in a standard diagnostic laboratory to detect microorganisms in blood culture material.
摘要
血培养常用于检测疑似血液感染患者体内的微生物。本研究评估了碱性洗涤/裂解程序，以提取临床血培养中微生物的 DNA。针对 16S rDNA（核糖体 DNA）基因和真菌 ITS（内部转录间隔区）基因的多重聚合酶链反应（PCR）被用作提取物中微生物 DNA 存在的可靠指标。共评估了 535 个 BacT/ALERT 阳性血培养瓶。多重 PCR 在 530 个 DNA 提取物中显示阳性结果，但 5 个 DNA 提取物结果为阴性。我们得出结论，碱性洗涤/裂解程序与多重 PCR 相结合是一种简单、敏感的方法，可在标准诊断实验室中用于检测血培养物中的微生物。
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J Microbiol Immunol Infect. 2007 Oct;40(5):445-9.
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