Comparative Analysis of Viral Concentration Methods for Detecting the HAV Genome Using Real-Time RT-PCR Amplification |
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| Authors: | Kang Bum Lee Hyeokjin Lee Sang-Do Ha Doo-Sung Cheon Changsun Choi |
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| Institution: | (1) Department of Food and Nutrition, School of Food Science and Technology, Chung-Ang University, 72-1, Nae-ri, Daeduck-myun, Ansung-si, Kyounggi-do, 456-756, South Korea;(2) Division of Hepatitis Virus, Korea Center for Disease Control and Prevention, Osong Health Technology Administration Complex, 187 Osongsaengmyeong2, Gangoe, Cheongwon, Chungcheongbuk-do, 363-951, South Korea;(3) School of Food Science and Technology, Chung-Ang University, 72-1, Nae-ri, Daeduck-myun, Ansung-si, Kyounggi-do, 456-756, South Korea; |
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| Abstract: | Hepatitis A is a major infectious disease epidemiologically associated with foodborne and waterborne outbreaks. Molecular detection using real-time RT-PCR to detect the hepatitis A virus (HAV) in contaminated vegetables can be hindered by low-virus recoveries during the concentration process and by natural PCR inhibitors in vegetables. This study evaluated three virus concentration methods from vegetables: polyethylene glycol (PEG) precipitation, ultrafiltration (UF), and immunomagnetic separation (IMS). UF was the most efficient concentration method, while PEG and IMS were very low for the recovery rate of HAV. These results demonstrate that UF is the most appropriate method for recovering HAV from contaminated vegetables and that this method combined with the real-time RT-PCR assay may be suitable for routine laboratory use. |
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