Cheung R C, Matsui S M, Greenberg H B
Department of Medicine, Palo Alto VA Medical Center, CA 94304.
J Clin Microbiol. 1994 Oct;32(10):2593-7. doi: 10.1128/jcm.32.10.2593-2597.1994.
We describe a rapid, sensitive, and economic method for detection of hepatitis C virus (HCV) RNA. This method uses silica particles for purification of nucleic acid and then a modified reverse transcription-PCR that minimizes the risk of contamination and reduces the amount of reagents used. We found purification by silica particles to be at least as sensitive and in certain circumstances more sensitive than that by traditional phenol-chloroform extraction. This improved sensitivity may be due to more efficient recovery of HCV RNA by silica particles. HCV RNA appears to bind to silica particles in a saturable fashion, and the addition of extraneous nucleic acids (salmon sperm DNA or tRNA) decreases the binding in a dose-related fashion. The reverse transcription-PCR is performed by using a modified single tube method which further simplifies and reduces the cost of this assay. Finally, this method may be applied to clinical specimens such as liver tissue.
我们描述了一种快速、灵敏且经济的丙型肝炎病毒（HCV）RNA检测方法。该方法使用硅胶颗粒纯化核酸，然后采用改良的逆转录-聚合酶链反应（RT-PCR），可将污染风险降至最低，并减少试剂用量。我们发现，硅胶颗粒纯化至少与传统酚-氯仿提取一样灵敏，在某些情况下甚至更灵敏。这种灵敏度的提高可能是由于硅胶颗粒能更有效地回收HCV RNA。HCV RNA似乎以饱和方式与硅胶颗粒结合，添加外源核酸（鲑鱼精DNA或tRNA）会以剂量相关方式降低结合。逆转录-聚合酶链反应采用改良的单管方法进行，进一步简化了该检测方法并降低了成本。最后，该方法可应用于肝组织等临床标本。
