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

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" method used currently in our laboratory. The best results were achieved with Total Quick RNA Cells & Tissues version mini (Talent) for RNA extraction and the Superscript(TM) One-Step RT-PCR System (Life Technologies) for the RT-PCR reaction, obtaining a detection limit of 0.1-1 pfu/mg of mussel tissue. A slightly lower sensitivity (in I log unit) 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 300 pfu/mg of tissue. Taken together, these findings indicate that the combination of Total Quick RNA Cells & Tissues version mini and Superscript(TM) 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. (C) 2003 Elsevier B.V. All rights reserved.