Detection of severe acute respiratory syndrome probable patients' virus RNA in Hangzhou by using a two loci and a modified nested real-time reverse transcription polymerase chain reaction / 中华预防医学杂志
Chinese Journal of Preventive Medicine
;
(12): 129-132, 2005.
Artigo
em Chinês
| WPRIM
| ID: wpr-282373
ABSTRACT
<p><b>OBJECTIVE</b>To detect the RNA of severe acute respiratory syndrome virus (SARS-CoV) by using reverse transcription polymerase chain reaction (RT-PCR) targeted for a two loci and a modified nested real-time RT-PCR as to improving the reliability and sensitivity of tests.</p><p><b>METHODS</b>A nested RT-PCR was used for detecting one fragment of SARS-CoV RNA in oropharyngeal swabs from 3 SARS probable patients, 4 SARS suspect patients and other 27 patients with fever in Hangzhou, and the nested RT-PCR product from one SARS probable patient was sequenced. Meanwhile in these 3 SARS probable patients, other three RT-PCR methods, including a hemi-nested RT-PCR targeted for another fragment of SARS-CoV RNA, a real-time RT-PCR and a modified nested real-time RT-PCR, were employed to detect SARS-CoV RNA.</p><p><b>RESULTS</b>Two positives were found in the 3 SARS probable patients, and none positive in 4 SARS suspect patients and other 27 patients with fever, using the nested RT-PCR. The sequence of the nested RT-PCR product from one SARS probable patient was identified with the counterpart of SARS-CoV genomes published in public database. The results of the hemi-nested RT-PCR, the real-time RT-PCR and the modified nested real-time RT-PCR in the 3 SARS patients were consistent with the one of the nested RT-PCR. During detecting specimen with low copies of RNA, a weak positive signal was produced after about 35 cycles in the real-time RT-PCR, but a strong positive signal was found only after 10 cycles in the modified nested real-time RT-PCR.</p><p><b>CONCLUSION</b>It might improve the reliability of test by employing RT-PCR targeted for two or more fragments in SARS-CoV genome. The modified nested real-time RT-PCR might have higher sensitivity than the routine real-time RT-PCR.</p>
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Virologia
/
RNA Viral
/
Sequência de Bases
/
Reprodutibilidade dos Testes
/
Sensibilidade e Especificidade
/
Reação em Cadeia da Polimerase Via Transcriptase Reversa
/
Síndrome Respiratória Aguda Grave
/
Coronavírus Relacionado à Síndrome Respiratória Aguda Grave
/
Diagnóstico
/
Genética
Tipo de estudo:
Estudo diagnóstico
/
Estudo prognóstico
Limite:
Adolescente
/
Adulto
/
Idoso
/
Aged80
/
Humanos
Idioma:
Chinês
Revista:
Chinese Journal of Preventive Medicine
Ano de publicação:
2005
Tipo de documento:
Artigo
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