Assessment of different commercial RNA-extraction and RT-PCR kits for detection of hepatitis A virus in mussel tissues.

摘要

In the present study, the efficiency of several nucleic acid extraction and RT-PCR commercial kits for the detection of hepatitis A virus (HAV) from seeded mussel tissue samples was evaluated in comparison with the in-house achieved with Total Quick RNA Cells & Tissues version mini (Talent) for RNA extraction and the Superscript One-Step RT-PCR System (Life Technologies) for the RT-PCR reaction, obtaining a detection limit of 0.1-1pfu/mg of mussel tissue. A slightly lower sensitivity (in 1logunit) was achieved using the Rneasy((R)) plant mini kit (Qiagen) and the Total Quick RNA Cells & Tissues version maxi in combination with the Superscript RT-PCR system. The conventional method usually employed in our laboratory resulted in a sensitivity of 300pfu/mg of tissue. Taken together, these findings indicate that the combination of Total Quick RNA Cells & Tissues version mini and Superscript One-Step RT-PCR System cannot only improve significantly the sensitivity for the HAV detection from mussel, but are also labor and time saving and easy to standardize.