Salmonella enterica subsp. enterica serovar Gallinarum (S. Gallinarum) is the causative agent of fowl typhoid, a major threat in poultry which causes mortality and high morbidity. The standard method for detection of S. Gallinarum relies on culture isolation and it requires more than five days for final confirmation. Recently, molecular methods have been developed, but they are not considered as golden standards, also because they do not allow pathogen quantification. Therefore, we developed a real time quantitative PCR (qPCR) which may represent a fast and accurate method that reduce drastically the extent of the analytical process, while being sensitive and specific. In order to grant specificity, a specific probe has been designed to detect the amplification products. The method has been tested in vitro and it has been proved to be effective for detection and absolute quantification of S. Gallinarum. In particular we found high sensitivity and specificity, with accuracy higher than 98%. Therefore, we find that the method may be suitable for diagnostic and other application, such as environmental monitoring.

Nuovo protocollo di qPCR per la diagnosi di Salmonella enterica serovar Gallinarum

Pugliese N;CIRCELLA, ELENA;DE VIRGILIO, Caterina;CAMARDA, Antonio
2015

Abstract

Salmonella enterica subsp. enterica serovar Gallinarum (S. Gallinarum) is the causative agent of fowl typhoid, a major threat in poultry which causes mortality and high morbidity. The standard method for detection of S. Gallinarum relies on culture isolation and it requires more than five days for final confirmation. Recently, molecular methods have been developed, but they are not considered as golden standards, also because they do not allow pathogen quantification. Therefore, we developed a real time quantitative PCR (qPCR) which may represent a fast and accurate method that reduce drastically the extent of the analytical process, while being sensitive and specific. In order to grant specificity, a specific probe has been designed to detect the amplification products. The method has been tested in vitro and it has been proved to be effective for detection and absolute quantification of S. Gallinarum. In particular we found high sensitivity and specificity, with accuracy higher than 98%. Therefore, we find that the method may be suitable for diagnostic and other application, such as environmental monitoring.
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11586/139577
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