Evaluation of the alkaline wash/lysis procedure for the molecular diagnosis of a positive bacterial blood culture in clinical routine practice
✍ Scribed by Sheng-Chuan Hsi; Jun-Ren Sun; Tzong-Shi Chiueh
- Publisher
- John Wiley and Sons
- Year
- 2010
- Tongue
- English
- Weight
- 104 KB
- Volume
- 24
- Category
- Article
- ISSN
- 0887-8013
No coin nor oath required. For personal study only.
✦ Synopsis
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 535BacT/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. J. Clin. Lab. Anal. 24:139–144, 2010. © 2010 Wiley‐Liss, Inc.