[Polymerase chain reaction (PCR) for the identification of toxigenic Vibrio cholerae O1 in oysters]. / La técnica de reacción de polimerización en cadena (PCR) para la identificación de Vibrio cholerae O1 toxigénico en ostiones.
Rev Latinoam Microbiol
; 36(4): 295-306, 1994.
Article
em Es
| MEDLINE
| ID: mdl-7701140
PCR was made with ctx2 (CGG GCA GAT TCT AGA CCT CCT G) y ctx3 (CGA TGA TCT TGG AGC ATT CCC AC) primers for subunit A of cholera toxin, 30 cycles of temperature on samples of 50 g of oysters added in 450 ml of peptone alcaline water that were inoculated with 15 x 10(6), 0.75 x 10(6) and 0.15 x 10(6) CFU/ml of toxigenic 6707 V. cholerae O1 reference strain. The samples were tested by three microbiological methods: INDRE's method uses 1 x 10(-1) dilution of sample, two fold pass to peptone alcaline water pH 9 incubated 18 h and 6 h at 37 degrees C, the Food and Drugs Administration (FDA) method uses 10(-1) to 10(-6) dilutions of sample, 6 h incubation and reincubation for 18 h at 37 and 42 degrees C and the Mexican laboratories (LMD) with 10(-4) to 10(-3) dilutions, the samples were incubated for 6 h and then reincubated for 18 h at two temperatures 37 and 42 degrees C. The PCR by INDRE's method was positive with 3 x 10(2) CFU/ml/g oyster. In the FDA's method the PCR detected DNA in 10(-4) dilution with 3 x 10(1) CFU/ml/g oyster and in LMD's method the PCR was positive in 10(-3) with 3 CFU/ml/g oyster. The results of the PCR were obtained between 5-6 h, and later V. cholerae O1 was isolated by three microbiological methods. The PCR reproducibility was better on DNA sample diluted 1:4 and 10 microliters of sample increased from 1:1000 to 1:10000 the sensitivity of PCR.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Ostreidae
/
Vibrio cholerae
/
DNA Bacteriano
/
Toxina da Cólera
/
Reação em Cadeia da Polimerase
Tipo de estudo:
Diagnostic_studies
Limite:
Animals
Idioma:
Es
Revista:
Rev Latinoam Microbiol
Ano de publicação:
1994
Tipo de documento:
Article
País de afiliação:
México
País de publicação:
México