deWit D, Wootton M, Allan B, Steyn L
Department of Medical Microbiology, Royal United Hospital, Bath, United Kingdom.
J Clin Microbiol. 1993 Aug;31(8):2204-7. doi: 10.1128/jcm.31.8.2204-2207.1993.
A simple method for the production of internal control DNA for two well-established Mycobacterium tuberculosis polymerase chain reaction assays is described. The internal controls were produced from Mycobacterium kansasii DNA with the same primers but at a lower annealing temperature than that used in the standard assays. In both assays, therefore, the internal control DNA has the same primer-binding sequences at the target DNA. One-microgram quantities of internal control DNA which was not contaminated with target DNA could easily be produced by this method. The inclusion of the internal control in the reaction mixture did not affect the efficiency of amplification of the target DNA. The method is simple and rapid and should be adaptable to most M. tuberculosis polymerase chain reaction assays.
本文描述了一种为两种成熟的结核分枝杆菌聚合酶链反应检测制备内部控制DNA的简单方法。内部控制DNA由堪萨斯分枝杆菌DNA使用相同引物制备,但退火温度低于标准检测中使用的温度。因此,在两种检测中,内部控制DNA在靶DNA处具有相同的引物结合序列。通过这种方法可以轻松制备出未被靶DNA污染的1微克量的内部控制DNA。将内部控制物加入反应混合物中不影响靶DNA的扩增效率。该方法简单快速,应适用于大多数结核分枝杆菌聚合酶链反应检测。