Establishment of a triplex real-time PCR for the detection of cholera toxin gene ctx and heat labile enterotoxin gene elt / 中华流行病学杂志
Chinese Journal of Epidemiology
;
(12): 720-723, 2014.
Artigo
em Chinês
| WPRIM
| ID: wpr-261648
ABSTRACT
<p><b>OBJECTIVE</b>To establish a triplex TaqMan real-time PCR system containing internal amplification control (IAC) to detect cholera toxin gene ctxA and enterotoxigenic Escherichia coli (ETEC)heat-labile enterotoxin gene elt.</p><p><b>METHODS</b>Primers and probes were designed based on the sequences of ctxA, elt and IAC. Both sensitivity and specificity were analyzed and interactions between different reactions were evaluated.</p><p><b>RESULTS</b>This system showed that the sensitivity of ctxA was 94 copies/reaction while the elt 79 copies/reaction and the amplification efficiency were 94.7% and 98.1%, respectively. Under the ratio of copy numbers on gene ctxA to elt as between 1 1-1 10, when both targets were detected, with impact was less on each other. However, when the amount of elt or ctxA was 100 times of IAC, the amplification of IAC was significantly inhibited.</p><p><b>CONCLUSION</b>This system showed both satisfactory sensitivity and specificity, thus could be used to detect pathogenic bacteria in diarrhea stools. The detection of IAC could prompt the presence of PCR inhibitors in samples being tested.</p>
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Toxina da Cólera
/
Sensibilidade e Especificidade
/
Enterotoxinas
/
Escherichia coli Enterotoxigênica
/
Reação em Cadeia da Polimerase em Tempo Real
/
Genética
/
Métodos
Tipo de estudo:
Estudo diagnóstico
Idioma:
Chinês
Revista:
Chinese Journal of Epidemiology
Ano de publicação:
2014
Tipo de documento:
Artigo
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