RESUMO
OBJECTIVE: To establish a simple rapid and sensitive nested RT-PCR method for detection of SARS coronavirus RNA by designing the specific primers for SARS and optimizing the parameters for PCR. METHODS: Primers and fluorescent probes were designed according to the sequences of SARS coronavirus genes available from GenBank. The optimization of the parameters for PCR was performed in PE 7700 thermal cycle. The 36 serum samples and 40 mouthwash of SARS patients and 80 samples of healthy people were tested. RESULTS: The positive rate of patient serum and mouthwash was 33.6%, (12/36) and 67.5%, (27/40), respectively, while the positive rate of healthy people was zero (0/160). CONCLUSION: The simple nested RT-PCR method was a rapid, efficient and sensitive one for SARS early diagnosis.
Assuntos
Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Síndrome Respiratória Aguda Grave/virologia , Coronavírus Relacionado à Síndrome Respiratória Aguda Grave/genética , Secreções Corporais/virologia , Primers do DNA , Humanos , RNA Viral/sangue , RNA Viral/genética , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Síndrome Respiratória Aguda Grave/sangue , Síndrome Respiratória Aguda Grave/diagnósticoRESUMO
OBJECTIVE: To develop a method for detection of coxsackie B virus type 1-6 by RT-PCR. METHODS: A pair of primers were designed to amplify all types of coxsackie B virus 1-6 efficiently. The PCR product was hybridized in micro-wells in which 6 type specific oligonucleotide probes had been coated respectively, colorimetric detection was performed to discriminate the types of coxsackie B virus. RESULTS: This method was shown to be concordant with the IgM ELISA, 71.7% of anti-coxsackie B positive cases could be detected by RT-PCR. CONCLUSION: The RT-PCR method can type coxsackie B virus efficiently and provides a tool for clinical diagnosis and epidemiological investigation.
