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Gene sequence analysis of SARS-associated coronavirus by nested RT-PCR



Gene sequence analysis of SARS-associated coronavirus by nested RT-PCR



Di 1 Jun Yi Da Xue Xue Bao 23(5): 421-423



To explore an effective means for the detection of Severe Acute Respiratory Syndrome (SARS)- associated coronavirus. 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. 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. 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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Accession: 049142389

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PMID: 12754117


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