Chandler L J, Borucki M K, Dobie D K, Wasieloski L P, Thompson W H, Gundersen C B, Case K, Beaty B J
Arthropod-borne and Infectious Diseases Laboratory, Department of Microbiology, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, Colorado 80523, USA.
J Clin Microbiol. 1998 Nov;36(11):3332-6. doi: 10.1128/JCM.36.11.3332-3336.1998.
A reverse transcription-PCR (RT-PCR) technique was used to detect La Crosse (LAC) virus RNA in the central nervous system (CNS) tissues of two patients who died of LAC encephalitis in 1960 and 1978. Viral RNA was readily detected by RT-PCR although the tissues had been stored frozen for up to 37 years. LAC virus was detected in the cerebral cortex but not in other CNS tissues. RT-PCR allowed detection of replicative forms of the virus, indicating that the virus was actively replicating in the specific CNS tissues. The small (S) RNA segments of the viruses from the CNS samples were demonstrated to be genetically similar by single-strand conformation polymorphism analyses. These S RNA segments were then sequenced; only two base changes were demonstrated between the 1960 and the 1978 samples, suggesting that LAC virus is genetically stable in areas of endemicity. The RT-PCR analyses of analyte directly from CNS tissues allows study of the virus without passage in cell culture.
采用逆转录聚合酶链反应(RT-PCR)技术检测了1960年和1978年死于拉克罗斯(LAC)脑炎的两名患者中枢神经系统(CNS)组织中的LAC病毒RNA。尽管这些组织已冷冻保存长达37年,但通过RT-PCR仍能轻松检测到病毒RNA。在大脑皮层中检测到了LAC病毒,但在其他CNS组织中未检测到。RT-PCR能够检测到病毒的复制形式,表明该病毒在特定的CNS组织中正在活跃复制。通过单链构象多态性分析表明,来自CNS样本的病毒的小(S)RNA片段在基因上相似。随后对这些S RNA片段进行了测序;1960年和1978年的样本之间仅显示出两个碱基变化,这表明LAC病毒在流行地区基因上是稳定的。直接对CNS组织中的分析物进行RT-PCR分析,无需在细胞培养中传代即可研究该病毒。