用于快速检测BK病毒的环介导等温扩增检测法的开发。
Development of a loop-mediated isothermal amplification assay for rapid detection of BK virus.
作者信息
Bista Bipin Raj, Ishwad Chandra, Wadowsky Robert M, Manna Pradip, Randhawa Parmjeet Singh, Gupta Gaurav, Adhikari Meena, Tyagi Rakhi, Gasper Gina, Vats Abhay
机构信息
Department of Pediatrics, Children's Hospital of Pittsburgh, 3705 Fifth Avenue, Pittsburgh, PA 15213, USA.
出版信息
J Clin Microbiol. 2007 May;45(5):1581-7. doi: 10.1128/JCM.01024-06. Epub 2007 Feb 21.
Loop-mediated isothermal amplification (LAMP) is a novel method for rapid amplification of DNA. Its advantages include rapidity and minimal equipment requirement. The LAMP assay was developed for BK virus (BKV), which is a leading cause of morbidity in renal transplant recipients. The characteristics of the assay, including its specificity and sensitivity, were evaluated. BKV LAMP was performed using various incubation times with a variety of specimens, including unprocessed urine and plasma samples. A ladder pattern on gel electrophoresis, typical of successful LAMP reactions, was observed specifically only for BKV and not for other viruses. The sensitivity of the assay with 1 h of incubation was 100 copies/tube of a cloned BKV fragment. Additionally, a positive reaction was visually ascertained by a simple color reaction using SYBR green dye. BKV LAMP was also successful for urine and plasma specimens without the need for DNA extraction. Due to its simplicity and specificity, the LAMP assay can potentially be developed for "point of care" screening of BKV.
环介导等温扩增技术（LAMP）是一种用于快速扩增DNA的新方法。其优点包括速度快和对设备要求极低。LAMP检测方法是针对BK病毒（BKV）开发的，BKV是肾移植受者发病的主要原因。对该检测方法的特性，包括其特异性和灵敏度进行了评估。使用各种孵育时间对包括未经处理的尿液和血浆样本在内的各种标本进行BKV LAMP检测。在凝胶电泳上观察到典型的成功LAMP反应的梯状条带，该条带仅在BKV检测中特异性出现，而在其他病毒检测中未出现。孵育1小时的检测方法灵敏度为每管100个克隆BKV片段拷贝。此外，使用SYBR绿色染料通过简单的颜色反应可直观确定阳性反应。BKV LAMP检测对尿液和血浆标本也很成功，无需进行DNA提取。由于其简单性和特异性，LAMP检测方法有可能用于BKV的“即时护理”筛查。
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