| 引用本文： |
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刘巍,古绍文,王树声,梁善凡,班华国,陈杰,孔健.用微量单管RT-PCR快速检测及定型脊髓灰质炎病毒[J].广西科学,1996,3(4):45-47,52. [点击复制]
- Liu Wei,Gu Shaowen,Wang Shusheng,Liang Shanfan,Ban Huaguo,Chen Jie,Kong Jian.Rapid Detecting and Identifying Poliovirus by Smaller Volume and One-Tube RT-PCR Assay[J].Guangxi Sciences,1996,3(4):45-47,52. [点击复制]
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| 摘要: |
| 介绍一种逆转录-聚合酶链反应(RT-PCR)的简单快速检测和定型脊髓灰质炎病毒(PV)的方法,无需提取PVRNA,共用一种缓冲系统,将RT和PCR试剂混合在20μL反应体系中,整个操作可置于一不间断温度循环程度程序中完成。一步法RT-PCR与常规RT-PCR及中和抗体试验对PV的检测结果完全一致,它的检测敏感度可达5TCID50/mL。该法需要的时间,费用和污染的都大大减少,而且特别适于临床常规诊断和鉴别PV的需要。 |
| 关键词: 脊髓灰质炎病毒 逆转录聚合酶链反应 一步法 检测 |
| DOI： |
| 投稿时间：1996-04-25修订日期：1996-09-20 |
| 基金项目: |
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| Rapid Detecting and Identifying Poliovirus by Smaller Volume and One-Tube RT-PCR Assay |
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Liu Wei1, Gu Shaowen1, Wang Shusheng1, Liang Shanfan1, Ban Huaguo1, Chen Jie1, Kong Jian2
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| (1.Guangxi Hygienic and Anti-epidemic Station, Taoyun Road, Nanning, Guangxi, 530021;2.National Vaccine and Serum Institute, Beijing, 100024) |
| Abstract: |
| We describe a simple method for the rapid detection and identification of poliovirus (PV) using one-step reverse transcription PCR (RT-PCR) method. Without RNA extraction, 20μL of mixed leagents for both RT and PCR were added into a tube,with sharing one buffer system. The whole procedure was performed with a single uninterrupted thermal cycling program. A good concordance was found among the PV neutralijation test, the routine RT-PCR and one-step RT-PCR. The sensitivity of one-step RT-PCR method reached the virus infectious titer of STCID50/mL. The needed time, cost and opportunity of contamination of this assay was reduced greatly. The one-step RT-PCR method is especially suitable for routine detection and identification of PV in clinical laboratories. |
| Key words: poliovirus RT-PCR one-step assay |
