Application of polymerase chain reaction assays to studies of herpes simplex virus latency.
Application of polymerase chain reaction assays to studies of herpes simplex virus latency.
复制标题
聚合酶链反应测定在单纯疱疹病毒潜伏期研究中的应用。
DOI：
10.1159/000150189
复制
发表时间：
1991
期刊：
影响因子：
4.6
通讯作者：
Openshaw,H
中科院分区：
文献类型：
--
作者：
Cantin,EM;Lange,W;Openshaw,H
We have amplified herpes simplex virus type 1 (HSV-1) DNA sequences from individual latently infected mouse trigeminal ganglia by polymerase chain reaction (PCR) assays. This report presents two useful modifications in the PCR technique. The first involves the use of two sets of closely spaced, oppositely oriented oligonucleotide primers and two rounds of 20–40 PCR cycles, first with the more widely spaced outer primers and then with the internal nested primers. This method enhanced the sensitivity of PCR detection as shown by assays of HSV-1 sequences in human brain. The second modification was designed to detect selectively HSV-1 sense or anti-sense RNA transcripts when both are present by adding a single primer during an initial reverse-transcriptase-mediated cDNA synthesis reaction. After destruction of the RNA template, standard PCR is initiated by the addition of the second primer and thermus aquaticus DNA polymerase (Taq). We show here applications of both of these modifications to amplify HSV-1 sequences from nervous system tissue.
