Duplex Real Time PCR for the detection of hepatitis A virus in shellfish using Feline Calicivirus as a process control.
J Virol Methods
; 163(1): 96-100, 2010 Jan.
Article
en En
| MEDLINE
| ID: mdl-19755130
ABSTRACT
The consumption of bivalve shellfish is a common cause of foodborne outbreaks of viral origin. The evaluation of the sanitary quality of these products, however, is still based on bacterial indicators of fecal contamination (Reg. (EC) No. 2073/2005 and No.1441/2007) even if it is known that they are not reliable indicators of viral contamination. In this study a duplex Real Time PCR method for quantitative detection of hepatitis A (HAV) in shellfish was developed. Feline Calicivirus (FCV) was used as a control for assessing the effectiveness of the concentration and extraction process and the ability to eliminate PCR inhibitors present in the food matrix. The specific primers and probes for detection of HAV and FCV, chosen respectively from the 5'-UTR region and in the ORF1 region, were labeled with two different dyes and detected simultaneously. The method was applied on spiked and non-spiked shellfish from a local market. The amplification of HAV in the presence of FCV showed good linearity (R(2)=0.994) and the sensitivity limit of the reaction was at least 5 x 10(2)TCID(50)g(-1) of an hepatopancreas extract.
Texto completo:
1
Base de datos:
MEDLINE
Asunto principal:
Reacción en Cadena de la Polimerasa de Transcriptasa Inversa
/
Virus de la Hepatitis A
/
Mytilus
/
Microbiología de Alimentos
Tipo de estudio:
Diagnostic_studies
Límite:
Animals
Idioma:
En
Revista:
J Virol Methods
Año:
2010
Tipo del documento:
Article
País de afiliación:
Italia