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KMID : 1007520080170030651
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Food Science and Biotechnology
2008 Volume.17 No. 3 p.651 ~ p.654
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Detection of Norovirus in Contaminated Ham by Reverse Transcriptase-PCR and Nested PCR
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Kim Seok-Ryel
Kim Du-Woon
Kwon Ki-Sung
Hwang In-Gyun
Oh Myung-Joo
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Abstract
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In order to enhance the efficacy of norovirus detection by reverse transcriptase-polymerase chain reaction (RTPCR) and nested PCR, this study developed a norovirus mRNA concentration method using poly oligo dT-conjugated magnetic beads. An efficient norovirus detection protocol was performed on commercial ham using 2 viral elution buffers (glycine buffer and Tris beef extract buffer) and 2 concentration solutions [polyethylene glycol (PEG) and zirconium hydroxide]. The different approaches were verified by RT-PCR and nested PCR. This method was performed on ham in less than 8 hr by artificial inoculation of serial dilutions of the virus ranging from 1,000 to 1 RT-PCR unit/mL. The viral extraction and concentration method had 10-fold higher sensitivity using the combination of Tris beef extract buffer and PEG as compared to glycine buffer and zirconium hydroxide. This method proved that RT-PCR and nested PCR have the sensitive ability to detect norovirus in commercial ham, in that norovirus was successfully detected in artificially contaminated samples at a detection level as low as 1-10 RT-PCR unit/mL. Overall, such a detection limit suggests this protocol is both quick and efficient in terms of its potential use for detecting norovirus in meat products.
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KEYWORD
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norovirus, reverse transcriptase-polymerase chain reaction, RT-PCR, ham, magnetic bead
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