用于直接分子检测血液中假马勒伯克霍尔德氏菌的 DNA 提取方法比较

Georgia Mataira, Fleur Francis, Jaimie Frazer, Robert Norton
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引用次数: 0

摘要

背景 Melioidosis 是由假马来伯克霍尔德氏菌(Burkholderia pseudomallei)引起的。从未经扩增的血液中直接进行分子检测仍然不敏感。方法 使用添加了乙二胺四乙酸的人体血液,试用了三种不同的提取方法--QIAamp UCP 病原体迷你试剂盒、高纯度 PCR 模板和 MagNA 纯病原体通用试剂盒。采用 III 型分泌系统 1 (TTSS-1) 聚合酶链反应进行检测。结果 QIAamp UCP 病原体迷你试剂盒性能最佳,检测限为 1.5×102 cfu/ml。结论 计划使用 QIAamp UCP 病原体迷你试剂盒对从类鼻疽患者采集的血液进行更大规模的研究。
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A comparison of DNA extraction methods used in the direct molecular detection of Burkholderia pseudomallei from blood
Background Melioidosis is caused by Burkholderia pseudomallei. Direct molecular detection from unamplified blood remains insensitive. Methods Three different extraction methods—QIAamp UCP Pathogen Mini Kit, High Pure PCR template and MagNA Pure Pathogen Universal—were trialled using spiked human ethylenediaminetetraacetic acid blood. A type III secretion system 1 (TTSS-1) polymerase chain reaction was used for detection. Results The QIAamp UCP Pathogen Mini Kit performed best, with a limit of detection of 1.5×102 cfu/ml. Conclusions It is planned to use the QIAamp UCP Pathogen Mini Kit to do a larger study on blood collected from patients with melioidosis.
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