A sensitive and reliable reverse transcriptase PCR-enzyme-linked immunosorbent assay for the detection of human pathogenic viruses in bivalve molluscs

Sarah Amelia Milne, S. Gallacher, P. Cash, D. N. Lees, K. Henshilwood, A. J. R. Porter

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

A colorimetric method, reverse transcriptase PCR with an enzyme-linked immunosorbent assay (RT-PCR-ELISA) was evaluated for ease of use, reliability, and sensitivity when detecting known human pathogenic virus present in shellfish, using a traditional polyethylene precipitation or immunocapture virus concentration method. The newly developed ELISA method could successfully detect enteroviruses and noroviruses in artificially and naturally contaminated shellfish. Overall, ELISA was shown to be a robust and sensitive method, which had a detection limit of 10 to 100 50% tissue culture infective dose enterovirus per gram of Crassostrea gigas (Pacific oyster) digestive gland and whole Mytilus edulis (common blue mussel). The technique was easily established in a new laboratory and required no specialized equipment. The method had a high sample throughput capable of screening 96 samples per run, making the technique extremely time efficient. RT-PCR-ELISA is a safe, quick, reliable technique, which has the potential for use as a standard virus detection method.

Original languageEnglish
Pages (from-to)1475-1482
Number of pages8
JournalJournal of Food Protection
Volume70
Issue number6
Publication statusPublished - Jun 2007

Keywords

  • hepatitis-A virus
  • polymerase-chain-reaction
  • round-structured viruses
  • norwalk-like viruses
  • time RT-PCR
  • human enteric viruses
  • shellfish tissues
  • nested-PCR
  • samples
  • enteroviruses

Cite this

A sensitive and reliable reverse transcriptase PCR-enzyme-linked immunosorbent assay for the detection of human pathogenic viruses in bivalve molluscs. / Milne, Sarah Amelia; Gallacher, S.; Cash, P.; Lees, D. N.; Henshilwood, K.; Porter, A. J. R.

In: Journal of Food Protection, Vol. 70, No. 6, 06.2007, p. 1475-1482.

Research output: Contribution to journalArticle

@article{55f1cfd881f248ee8d88847a510d4fc8,
title = "A sensitive and reliable reverse transcriptase PCR-enzyme-linked immunosorbent assay for the detection of human pathogenic viruses in bivalve molluscs",
abstract = "A colorimetric method, reverse transcriptase PCR with an enzyme-linked immunosorbent assay (RT-PCR-ELISA) was evaluated for ease of use, reliability, and sensitivity when detecting known human pathogenic virus present in shellfish, using a traditional polyethylene precipitation or immunocapture virus concentration method. The newly developed ELISA method could successfully detect enteroviruses and noroviruses in artificially and naturally contaminated shellfish. Overall, ELISA was shown to be a robust and sensitive method, which had a detection limit of 10 to 100 50{\%} tissue culture infective dose enterovirus per gram of Crassostrea gigas (Pacific oyster) digestive gland and whole Mytilus edulis (common blue mussel). The technique was easily established in a new laboratory and required no specialized equipment. The method had a high sample throughput capable of screening 96 samples per run, making the technique extremely time efficient. RT-PCR-ELISA is a safe, quick, reliable technique, which has the potential for use as a standard virus detection method.",
keywords = "hepatitis-A virus, polymerase-chain-reaction, round-structured viruses, norwalk-like viruses, time RT-PCR, human enteric viruses, shellfish tissues, nested-PCR, samples, enteroviruses",
author = "Milne, {Sarah Amelia} and S. Gallacher and P. Cash and Lees, {D. N.} and K. Henshilwood and Porter, {A. J. R.}",
year = "2007",
month = "6",
language = "English",
volume = "70",
pages = "1475--1482",
journal = "Journal of Food Protection",
issn = "0362-028X",
publisher = "International Association for Food Protection",
number = "6",

}

TY - JOUR

T1 - A sensitive and reliable reverse transcriptase PCR-enzyme-linked immunosorbent assay for the detection of human pathogenic viruses in bivalve molluscs

AU - Milne, Sarah Amelia

AU - Gallacher, S.

AU - Cash, P.

AU - Lees, D. N.

AU - Henshilwood, K.

AU - Porter, A. J. R.

PY - 2007/6

Y1 - 2007/6

N2 - A colorimetric method, reverse transcriptase PCR with an enzyme-linked immunosorbent assay (RT-PCR-ELISA) was evaluated for ease of use, reliability, and sensitivity when detecting known human pathogenic virus present in shellfish, using a traditional polyethylene precipitation or immunocapture virus concentration method. The newly developed ELISA method could successfully detect enteroviruses and noroviruses in artificially and naturally contaminated shellfish. Overall, ELISA was shown to be a robust and sensitive method, which had a detection limit of 10 to 100 50% tissue culture infective dose enterovirus per gram of Crassostrea gigas (Pacific oyster) digestive gland and whole Mytilus edulis (common blue mussel). The technique was easily established in a new laboratory and required no specialized equipment. The method had a high sample throughput capable of screening 96 samples per run, making the technique extremely time efficient. RT-PCR-ELISA is a safe, quick, reliable technique, which has the potential for use as a standard virus detection method.

AB - A colorimetric method, reverse transcriptase PCR with an enzyme-linked immunosorbent assay (RT-PCR-ELISA) was evaluated for ease of use, reliability, and sensitivity when detecting known human pathogenic virus present in shellfish, using a traditional polyethylene precipitation or immunocapture virus concentration method. The newly developed ELISA method could successfully detect enteroviruses and noroviruses in artificially and naturally contaminated shellfish. Overall, ELISA was shown to be a robust and sensitive method, which had a detection limit of 10 to 100 50% tissue culture infective dose enterovirus per gram of Crassostrea gigas (Pacific oyster) digestive gland and whole Mytilus edulis (common blue mussel). The technique was easily established in a new laboratory and required no specialized equipment. The method had a high sample throughput capable of screening 96 samples per run, making the technique extremely time efficient. RT-PCR-ELISA is a safe, quick, reliable technique, which has the potential for use as a standard virus detection method.

KW - hepatitis-A virus

KW - polymerase-chain-reaction

KW - round-structured viruses

KW - norwalk-like viruses

KW - time RT-PCR

KW - human enteric viruses

KW - shellfish tissues

KW - nested-PCR

KW - samples

KW - enteroviruses

M3 - Article

VL - 70

SP - 1475

EP - 1482

JO - Journal of Food Protection

JF - Journal of Food Protection

SN - 0362-028X

IS - 6

ER -