Use of magnetic beads for tissue DNA extraction and IS6110 Mycobacterium tuberculosis PCR
Use of magnetic beads for tissue DNA extraction and IS6110 Mycobacterium tuberculosis PCR
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DOI：
10.1136/mp.52.3.158
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发表时间：
1999-06-01
期刊：
影响因子：
--
通讯作者：
Costanzi, G
中科院分区：
文献类型：
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作者：
Caldarelli-Stefano, R;Vago, L;Costanzi, G
聚合酶链反应（PCR）技术越来越多地用于诊断结核分枝杆菌感染，并可用于从冷冻和福尔马林固定的石蜡包埋组织中获得的DNA。然而，通过常规的苯酚/氯仿方法提取DNA是耗时的，并且需要使用潜在危险的化学试剂。本文描述了一种基于使用磁珠从常规福尔马林固定、石蜡包埋的组织和冷冻组织中提取结核分枝杆菌DNA的方法。从人类免疫缺陷病毒（HIV）和结核病患者的脑、淋巴结和肺组织中提取DNA，并与酚/氯仿法进行比较。磁珠提取过程需要不到两个小时，包括组织切片脱蜡所需的时间。在所有情况下，用两种方法提取的DNA成功地通过PCR扩增结核分枝杆菌IS 6110序列。磁珠DNA提取可用于冷冻和存档组织：该方法可靠，简单，灵敏，快速;此外，它不使用危险的程序或专门的实验室设备，可用于从各种人体组织中常规分离DNA。
Polymerase chain reaction (PCR) techniques are used increasingly for the diagnosis of Mycobacterium tuberculosis infection and can be used on the DNA obtained from both frozen and formalin fixed, paraffin wax embedded tissues. However, the extraction of DNA by means of the conventional phenol/chloroform method is time consuming and requires the use of potentially dangerous chemical reagents. This paper describes a method based upon the use of magnetic beads for the extraction of M tuberculosis DNA from both routinely formalin fixed, paraffin wax embedded tissues and frozen tissues. Magnetic bead extracted DNA from brain, lymph node, and lung tissues collected from patients with human immunodeficiency virus and tuberculosis was compared with that extracted using the phenol/chloroform method. The magnetic bead extraction procedure requires less than two hours, including the time necessary to dewax the tissue sections. In all cases, the DNA extracted with both methods was amplified successfully by PCR for the M tuberculosis IS6110 sequence. Magnetic bead DNA extraction can be used on both frozen and archival tissues: the method is reliable, simple, sensitive, and rapid; in addition, it does not use hazardous procedures or specialised laboratory equipment and can be used for routine DNA isolation from various human tissues.