Establishment of a fluorescent real-time quantitative RT-PCR assay for detection of genotype 4 hepatitis E virus in swine stools / 病毒学报
Chinese Journal of Virology
;
(6): 33-39, 2010.
Artigo
em Chinês
| WPRIM
| ID: wpr-297921
ABSTRACT
The primers and probes for the Real-time RT-PCR were designed based on the multiple sequence (swine and humans HEV strains) alignments of the ORF3 region of genotype 4 HEV. A rapid, sensitive and stable TaqMan Real-time RT-PCR assay was established, and its specificity and sensitivity were assessed, and comparison of the Real-time RT-PCR with conventional and nested RT-PCR was performed. The results found that the crossing points showed linearly proportional to the logarithm of the input copy number. The correlation coefficient (R2) and the slope value of the standard curves with plasmid DNA were 0.994 and -3.312, respectively. The efficiency (E) of the PCR was 100%. Coefficients of variation values of the different diluted plasmid DNA were low in the same or different repeated experimental group. In addition, the assay was able to correctly detect genotype 4 HEV RNA from swine fecal samples. The sensitivity of established assay was 100-fold higher than that of conventional RT-PCR and 10-fold higher than nested RT-PCR.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Suínos
/
Virologia
/
Reservatórios de Doenças
/
Sensibilidade e Especificidade
/
Classificação
/
Vírus da Hepatite E
/
Hepatite E
/
Primers do DNA
/
Reação em Cadeia da Polimerase Via Transcriptase Reversa
/
Fezes
Tipo de estudo:
Estudo diagnóstico
Limite:
Animais
/
Humanos
Idioma:
Chinês
Revista:
Chinese Journal of Virology
Ano de publicação:
2010
Tipo de documento:
Artigo
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