A simple and sensitive method to extract bacterial, yeast and fungal DNA from blood culture material

被引：125

作者：

Miller, BC

Jiru, X

Moore, JE ^{[1
]}

Earle, JAP

机构：

[1] Belfast City Hosp, Dept Bacteriol, No Ireland Publ Hlth Lab, Belfast BT9 7AD, Antrim, North Ireland

[2] Queens Univ Belfast, Sch Biol & Biochem, Belfast BT7 1NN, Antrim, North Ireland

来源：

JOURNAL OF MICROBIOLOGICAL METHODS | 2000年 / 42卷 / 02期

关键词：

bacteria; blood-culture; contaminant; DNA extraction; fungi; PCR inhibitor;

D O I：

10.1016/S0167-7012(00)00174-3

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

This study investigated the various commercially available kits and 'in-house' methods to extract DNA from Gramnegative and Gram-positive bacteria, yeast and fungal agents in commonly employed blood culture material. The main methods investigated were as follows; Qiagen QIAmp Blood kit, Roche high PCR template preparation kit, Puregene DNA extraction kit, boiling, glass beads/sonication and wash/alkali/heat lysis. The results indicated that a simple wash/alkali/heat lysis method was the most sensitive, reproducible, simple and cost-effective extraction method. This was the only method which removed any PCR inhibitors and inherent DNA which existed in virgin BacT/Alert aerobic, anaerobic and paediatric blood culture material. Contaminating microbial DNA from Lactococcus lactis or Bacillus coagulans was identified in all batches of BacT/Alert(R) FAN(R) aerobic blood culture material examined. (C) 2000 Elsevier Science B.V. All rights reserved.

引用

页码：139 / 147

页数：9

共 12 条

[1] Detection and identification of fungal pathogens in blood by using molecular probes [J].