title: Evaluation of the alkaline wash/lysis procedure for the molecular diagnosis of a positive bacterial blood culture in clinical routine practice.
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.
authors.affiliation: Hsin Sheng College of Medical Care and Management, Taoyuan, Taiwan.
authors.forename: Sheng-Chuan
authors.initials: SC
authors.lastname: Hsi
authors.forename: Jun-Ren
authors.initials: JR
authors.lastname: Sun
authors.forename: Tzong-Shi
authors.initials: TS
authors.lastname: Chiueh
mesh_terms: D016470:Bacteremia / Q000097:blood / Q000175:diagnosis*; D001769:Blood / Q000382:microbiology*; D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics / Q000302:isolation & purification*; D004271:DNA, Fungal / Q000032:analysis / Q000235:genetics / Q000302:isolation & purification; D021901:DNA, Intergenic / Q000235:genetics; D016469:Fungemia / Q000175:diagnosis; D006801:Humans; D058889:Molecular Typing / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012680:Sensitivity and Specificity
chemical_list: D004269:DNA, Bacterial; D004271:DNA, Fungal; D021901:DNA, Intergenic; D012336:RNA, Ribosomal, 16S