IMPROVED DETECTION OF VIRUSES BY ELECTRON-MICROSCOPY AFTER DIRECT ULTRA-CENTRIFUGE PREPARATION OF SPECIMENS
IMPROVED DETECTION OF VIRUSES BY ELECTRON-MICROSCOPY AFTER DIRECT ULTRA-CENTRIFUGE PREPARATION OF SPECIMENS
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DOI：
10.1128/jcm.14.2.210-221.1981
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发表时间：
1981-01-01
影响因子：
9.4
通讯作者：
KLISKO, B
中科院分区：
文献类型：
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作者：
HAMMOND, GW;HAZELTON, PR;KLISKO, B
Beckman Airfuge 空气涡轮超速离心机和新型 EM-90 颗粒计数转子经过改造，可改进临床样本中病毒的 EM 检测。样品通过离心澄清，在 EM-90 转子中直接沉淀到 Formvar 涂层的铜网上，并用 1.5% 磷钨酸钠染色。脊髓灰质炎病毒、SA11轮状病毒、单纯疱疹病毒和痘苗病毒的部分纯化制剂的系列稀释液的病毒计数和终点滴定显示增加了.apprx。比相同材料的未浓缩制剂的病毒滴度高 1.5 至 3.0 log10。与不浓缩病毒的方法相比，通过直接超速离心制备临床样品后，轮状病毒阳性标本的产量增加了 14%（82 比 72）。一项前瞻性研究表明，通过直接超速离心制备标本时，腺病毒、疱疹病毒和肠道病毒的检出率会增加。 Airfuge 超速离心机中使用 EM-90 转子进行直接超速离心是一种快速浓缩方法，可提高通过 EM 从临床标本中检测病毒的速率和产量，并且易于适用于诊断病毒学实验室。
The Beckman Airfuge air turbine ultracentrifuge and the new EM-90 particle-counting rotor were adapted to improve detection by EM of viruses in clinical specimens. Samples were clarified by centrifugation, pelleted in the EM-90 rotor directly to Formvar-coated Cu grids and stained with 1.5% sodium phosphotungstate. Virus counts and endpoint titrations of serial dilutions of partially purified preparations of poliovirus, SA11 rotavirus, herpes simplex virus and vaccinia virus showed an increase of .apprx. 1.5 to 3.0 log10 over the virus titers of unconcentrated preparations of the same material. An increased yield of 14% more positive specimens for rotavirus was obtained after preparation of clinical samples by direct ultracentrifugation vs. a method without virus concentration (82 vs. 72). A prospective study showed that detection of adenoviruses, herpesviruses and enteroviruses increased when specimens were prepared by direct ultracentrifugation. Direct ultracentrifugation with the EM-90 rotor in the Airfuge ultracentrifuge is a rapid concentration method which enhances the rate and yield of virus detection from clinical specimens by EM and is easily adaptable to a diagnostic virology laboratory.