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Rapid detection method for hepatitis A virus from lettuce by a combination of filtration and integrated cell culture-real-time reverse transcription PCR

Hyeon, Ji-Yeon ; Chon, Jung-Whan ; Park, Chankyu ; Lee, Joong-Bok ; Choi, In-Soo ; Kim, Moo-Sang ; Seo, Kun-Ho

Journal of food protection, October 2011, Vol.74(10), pp.1756-61 [Peer Reviewed Journal]

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  • Title:
    Rapid detection method for hepatitis A virus from lettuce by a combination of filtration and integrated cell culture-real-time reverse transcription PCR
  • Author: Hyeon, Ji-Yeon ; Chon, Jung-Whan ; Park, Chankyu ; Lee, Joong-Bok ; Choi, In-Soo ; Kim, Moo-Sang ; Seo, Kun-Ho
  • Description: We have developed a rapid and simple method for filtration using a positively charged membrane to concentrate hepatitis A virus (HAV) from lettuce and an integrated cell culture-real-time reverse transcription PCR (ICC-real-time RT-PCR) to detect infectious HAV. The most suitable buffer for HAV concentration by filtration was 100 mM Tris-HCl, 50 mM glycine (pH 9.5). Filtration using the NanoCeram matrix was compared with polyethylene glycol precipitation for viral concentration from lettuce inoculated with 6 log RNA copies of HAV. The recovery rate of filtration was statistically higher than that of polyethylene glycol precipitation (47.3 versus 24.9%, respectively). The sensitivity of ICC-real-time RT-PCR for detection of infectious HAV was determined by inoculation of FRhK-4 cells with HAV (4 log to 0 log RNA copies). ICC-real-time RT-PCR detected infectious HAV on average 5 days earlier than cytopathic effects at all inoculation levels. HAV recovered from lettuce (approximately 3 log RNA copies) was also analyzed with ICC-real-time RT-PCR. Infectious HAV was detected within 2 days postinfection by ICC-real-time RT-PCR, whereas cytopathic effects were not observed until 7 days postinfection. Coupled with a virus concentration and purification system using a positively charged membrane, ICC-real-time RT-PCR has the potential to become a novel and rapid method for the detection of infectious HAV in vegetables.
  • Is Part Of: Journal of food protection, October 2011, Vol.74(10), pp.1756-61
  • Identifier: E-ISSN: 1944-9097 ; PMID: 22004827 Version:1 ; DOI: 10.4315/0362-028X.JFP-11-155
  • Subjects: Filtration -- Methods ; Food Contamination -- Analysis ; Hepatitis A Virus -- Isolation & Purification ; Lettuce -- Virology ; Reverse Transcriptase Polymerase Chain Reaction -- Methods
  • Language: English
  • Source: MEDLINE/PubMed (U.S. National Library of Medicine)

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