Author: Wang, Yan; Ma, Wen-li; Song, Yan-bin; Xiao, Wei-wei; Zhang, Bao; Huang, Hai; Wang, Hong-min; Ma, Xiao-dong; Zheng, Wen-ling
Title: [Gene sequence analysis of SARS-associated coronavirus by nested RT-PCR]. Cord-id: 03h6xkig Document date: 2003_1_1
ID: 03h6xkig
Snippet: OBJECTIVE To explore an effective means for the detection of Severe Acute Respiratory Syndrome (SARS)- associated coronavirus. METHODS The RNAs of the virus contained in the sputum samples from established SARS patients were extracted and reversely transcripted, followed by nested PCR using the reversely transcripted cDNA as the template. The PCR products were cloned then into the pMD18-T vectors, followed by sequence analysis. RESULTS Specific fragments were amplified from the sputum samples of
Document: OBJECTIVE To explore an effective means for the detection of Severe Acute Respiratory Syndrome (SARS)- associated coronavirus. METHODS The RNAs of the virus contained in the sputum samples from established SARS patients were extracted and reversely transcripted, followed by nested PCR using the reversely transcripted cDNA as the template. The PCR products were cloned then into the pMD18-T vectors, followed by sequence analysis. RESULTS Specific fragments were amplified from the sputum samples of SARS patients, which were confirmed by DNA cloning and sequencing to belong to SARS-associated coronavirus. The Result of Blast shows only the difference in one nucleic acid from the TOR2 strain of SARS-associated coronavirus. CONCLUSION Sequence analysis has confirmed the existence of SARS-associated coronavirus in the sputum samples of SARS patients, and nested RT-PCR is a quick, easy, and convenient way for the detection of the virus.
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