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. 2010 May 13;24(3):139–144. doi: 10.1002/jcla.20396

Evaluation of the alkaline wash/lysis procedure for the molecular diagnosis of a positive bacterial blood culture in clinical routine practice

Sheng‐Chuan Hsi 1,2, Jun‐Ren Sun 3,†, Tzong‐Shi Chiueh 3,†,✉
PMCID: PMC6647685  PMID: 20486192

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.

Keywords: blood culture, alkaline wash, lysis procedure

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