Richardson K J, Margolin A B, Gerba C P
Department of Microbiology and Immunology, University of Arizona, Tucson 85721.
J Virol Methods. 1988 Oct;22(1):13-21. doi: 10.1016/0166-0934(88)90083-3.
Rapid and sensitive methods are needed for the detection of enteric viruses to ensure proper drinking water quality. Gene probes have been shown to be useful for this purpose. Previously, samples to be assayed were treated with a series of phenol-chloroform extractions to release the viral nucleic acid. We have developed a more rapid procedure for liberating or exposing the genome of poliovirus for probing. In this study, a poliovirus model was used to test the ability of heat (65 degrees C for 30 min) for release or exposure of viral nucleic acid. Several different RNase inhibitors were tested for their ability to prevent viral RNA degradation. A comparison of the two methods indicates phenol-chloroform extraction is not necessary before probing. In addition to saving 2-4 h of time, maximum sensitivity levels were consistently obtained using this novel procedure.
为确保饮用水质量，需要快速灵敏的方法来检测肠道病毒。基因探针已被证明可用于此目的。以前，待检测的样本要用一系列苯酚 - 氯仿萃取法来释放病毒核酸。我们已开发出一种更快速的程序来释放或暴露脊髓灰质炎病毒的基因组以便进行探测。在本研究中，使用脊髓灰质炎病毒模型来测试加热（65摄氏度30分钟）释放或暴露病毒核酸的能力。测试了几种不同的核糖核酸酶抑制剂防止病毒RNA降解的能力。两种方法的比较表明，探测前无需进行苯酚 - 氯仿萃取。除了节省2 - 4小时时间外，使用这种新程序始终能获得最高灵敏度水平。
