The efficiency of different commercial RNA extraction kits for the detection of hepatitis A virus (HAV) from seeded strawberry samples was assessed by standard RT-PCR and real time RT-PCR (RT-qPCR). The best results with standard RT-PCR were achieved with Aurum Total RNA extraction kit (BioRad), obtaining a detection limit of 5e6.25 pfu/mg of tissue. A slightly lower sensitivity was rendered by the RNeasy Plant mini kit (Qiagen) (10e12.5 pfu/mg tissue), while the Total Quick RNA Cells and Tissues kit version mini (Talent) rendered a detection limit of 50e100 pfu/mg of tissue. The other tested commercial kits showed worse detection limits (>500 pfu/mg). With RT-qPCR and ten fold diluted RNA all the kits showed an increase of sensitivity, detecting the kits from Qiagen, Talent and BioRad down to 0.05 pfu/mg of strawberry homogenate. These findings indicate that the use of Aurum Total RNA extraction kit, with standard RT-PCR technique or RT-qPCR, can not only be labor and time saving but also helpful to improve the sensitivity for the HAV detection from fruits and to facilitate the standardization of detection methods among laboratories.

Evaluation of different RNA-extraction kits for sensitive detection of Hepatitis A virus in strawberry samples / S. Bianchi, A. Dal Vecchio, M.L. Vilariño, J. Lopez Romalde. - In: FOOD MICROBIOLOGY. - ISSN 0740-0020. - 28:1(2011), pp. 38-42. [10.1016/j.fm.2010.08.002]

Evaluation of different RNA-extraction kits for sensitive detection of Hepatitis A virus in strawberry samples

S. Bianchi
Primo
;
2011

Abstract

The efficiency of different commercial RNA extraction kits for the detection of hepatitis A virus (HAV) from seeded strawberry samples was assessed by standard RT-PCR and real time RT-PCR (RT-qPCR). The best results with standard RT-PCR were achieved with Aurum Total RNA extraction kit (BioRad), obtaining a detection limit of 5e6.25 pfu/mg of tissue. A slightly lower sensitivity was rendered by the RNeasy Plant mini kit (Qiagen) (10e12.5 pfu/mg tissue), while the Total Quick RNA Cells and Tissues kit version mini (Talent) rendered a detection limit of 50e100 pfu/mg of tissue. The other tested commercial kits showed worse detection limits (>500 pfu/mg). With RT-qPCR and ten fold diluted RNA all the kits showed an increase of sensitivity, detecting the kits from Qiagen, Talent and BioRad down to 0.05 pfu/mg of strawberry homogenate. These findings indicate that the use of Aurum Total RNA extraction kit, with standard RT-PCR technique or RT-qPCR, can not only be labor and time saving but also helpful to improve the sensitivity for the HAV detection from fruits and to facilitate the standardization of detection methods among laboratories.
Hepatitis A virus; Fruits; Detection; Commercial RNA extraction kits
Settore MED/42 - Igiene Generale e Applicata
Settore AGR/15 - Scienze e Tecnologie Alimentari
Settore MED/07 - Microbiologia e Microbiologia Clinica
14-ago-2010
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/2434/161353
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