| PCR条件对扩增SARS-CoV多聚酶部分基因的影响 |
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| 引用本文： | 赵卫,晏辉钧,张文炳,方丹云,周经姣,胡琼,龙北国,江丽芳.PCR条件对扩增SARS-CoV多聚酶部分基因的影响[J].生物技术,2003,13(6):11-12. |
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| 作者姓名： | 赵卫 晏辉钧 张文炳 方丹云 周经姣 胡琼 龙北国 江丽芳 |
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| 作者单位： | 1. 第一军医大学热带军队卫生学系微生物学教研室,广东,广州,510515
2. 广州中山大学中山医学院微生物学教研室,广东,广州,510080 |
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| 基金项目： | 广东省科技攻关项目 (“SARS病毒感染细胞模型的建立及初步应用”) |
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| 摘 要： | 目的：对该实验室已建立的检测SARS冠状病毒多聚酶基因的套式RT-PCR方法进行优化。方法：从SAPS病人的嗽口水标本中提取RNA，调整套式PCR的退火温度，扩增SARS冠状病毒多聚酶部分基因。扩增出的阳性片段连接入pGEM-T载体中，测序后比较其与已知SARS冠状病毒的同源性。结果：通过改变PCR条件，成功从一SARS病人的嗽口水中扩增出SARS冠状病毒多聚酶部分基因。结论：针对不同标本优化PCR反应条件非常重要。
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| 关 键 词： | PCR条件 SARS冠状病毒 套式RT-PCR 退火温度 非典型肺炎 |
| 文章编号： | 1004-311X(2003)06-0011-02 |
| 修稿时间： | 2003年7月5日 |
Effects of PCR Conditions on the Amplification of the Partial Polymerase Gene of SARS-CoV |
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| ZHAO Wei,YAN Hui-jun,ZHANG Wen-bing,FANG Dan-yun,ZHOU Jing-jiao,HU Qiong,LONG Bei-guo.Effects of PCR Conditions on the Amplification of the Partial Polymerase Gene of SARS-CoV[J].Biotechnology,2003,13(6):11-12. |
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| Authors: | ZHAO Wei YAN Hui-jun ZHANG Wen-bing FANG Dan-yun ZHOU Jing-jiao HU Qiong LONG Bei-guo |
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| Institution: | ZHAO Wei~1,YAN Hui-jun~2,ZHANG Wen-bing~1,FANG Dan-yun~2,ZHOU Jing-jiao~2,HU Qiong~1,LONG Bei-guo~ |
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| Abstract: | Objective:To optimize the RT-PCR method for detection the partial polymerase gene of SARS coronavirus (SARS-CoV) in our laboratory.Methods:The RNAs of the SARS-CoV contained in the gargles patients with SARS were extracted.The annealing temperatures were modified and then the partial polymerase gene was amplified.The amplicons were cloned into the pGEM-T vectors and then sequenced.The obtained sequences were compared with the published sequence of SARS-CoV.Results:The partial polymerase gene was got from the gargles of a patients with SARS after the annealing temperatures were changed.Conclusion It is very important to optimize PCR conditions according to different samples of SARS patients. |
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| Keywords: | SARS coronavirus (SARS-CoV) Nested RT-PCR Detection |
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