Simple and Rapid Detection of Human Norovirus from Produce Using SYBR Green I-based Real-time RT-PCR
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2011
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Several foodborne norovirus gastroenteritis outbreaks have been linked to fresh produce. Rapid and sensitive detection can help prevent the release of contaminated produce items in the market. The objectives of this study were to apply a relatively inexpensive SYBR Green I-based real-time RT-PCR assay for the rapid detection of human norovirus (NoV) GI and GII on the surfaces of lettuce, cherry tomatoes, and green onions. Each washed produce commodity (25 g) was spiked with serial dilutions of NoV GI and GII stool samples. RNA was eluted from the produce surface and extracted using the TRIzol((TM)) method. This was followed by detection using SYBR Green I real-time RT-PCR with primers specific for NoV GI (COG1F-COG1R) and GII (COG2F-COG2R) along with an internal RNA amplification control. End-point detection limits from lettuce and tomatoes were found to be 10 RT-PCR units/25 g for GI and GII and 1 RT-PCR unit/25 g for GI and 10 RT-PCR units/25 g for GII from green onions. These result...s were confirmed by Tm analysis (showing peaks at 81.5 and 84A degrees C for GI and GII, respectively; and 83A degrees C for the IAC) as well as agarose gel electrophoresis that confirmed products of similar to 95 bp for GI and GII and similar to 155 bp for the RNA IAC. Results could be obtained within one working day, showing potential for routine use in diagnostics and monitoring of NoV contamination by the produce industry.
Ključne reči:
Fresh produce / Noroviruses / Detection / SYBR Green I real-time RT-PCRIzvor:
Food and Environmental Virology, 2011, 3, 3-4, 121-129Izdavač:
- Springer, New York
Finansiranje / projekti:
- TN Agricultural Experiment Station (TEN) [00391]
- Fulbright Program [68432926]
DOI: 10.1007/s12560-011-9066-5
ISSN: 1867-0334
WoS: 000297228000004
Scopus: 2-s2.0-81355161363
Institucija/grupa
Poljoprivredni fakultetTY - JOUR AU - Radin, Dragoslava AU - D'Souza, Doris H. PY - 2011 UR - http://aspace.agrif.bg.ac.rs/handle/123456789/2714 AB - Several foodborne norovirus gastroenteritis outbreaks have been linked to fresh produce. Rapid and sensitive detection can help prevent the release of contaminated produce items in the market. The objectives of this study were to apply a relatively inexpensive SYBR Green I-based real-time RT-PCR assay for the rapid detection of human norovirus (NoV) GI and GII on the surfaces of lettuce, cherry tomatoes, and green onions. Each washed produce commodity (25 g) was spiked with serial dilutions of NoV GI and GII stool samples. RNA was eluted from the produce surface and extracted using the TRIzol((TM)) method. This was followed by detection using SYBR Green I real-time RT-PCR with primers specific for NoV GI (COG1F-COG1R) and GII (COG2F-COG2R) along with an internal RNA amplification control. End-point detection limits from lettuce and tomatoes were found to be 10 RT-PCR units/25 g for GI and GII and 1 RT-PCR unit/25 g for GI and 10 RT-PCR units/25 g for GII from green onions. These results were confirmed by Tm analysis (showing peaks at 81.5 and 84A degrees C for GI and GII, respectively; and 83A degrees C for the IAC) as well as agarose gel electrophoresis that confirmed products of similar to 95 bp for GI and GII and similar to 155 bp for the RNA IAC. Results could be obtained within one working day, showing potential for routine use in diagnostics and monitoring of NoV contamination by the produce industry. PB - Springer, New York T2 - Food and Environmental Virology T1 - Simple and Rapid Detection of Human Norovirus from Produce Using SYBR Green I-based Real-time RT-PCR EP - 129 IS - 3-4 SP - 121 VL - 3 DO - 10.1007/s12560-011-9066-5 ER -
@article{ author = "Radin, Dragoslava and D'Souza, Doris H.", year = "2011", abstract = "Several foodborne norovirus gastroenteritis outbreaks have been linked to fresh produce. Rapid and sensitive detection can help prevent the release of contaminated produce items in the market. The objectives of this study were to apply a relatively inexpensive SYBR Green I-based real-time RT-PCR assay for the rapid detection of human norovirus (NoV) GI and GII on the surfaces of lettuce, cherry tomatoes, and green onions. Each washed produce commodity (25 g) was spiked with serial dilutions of NoV GI and GII stool samples. RNA was eluted from the produce surface and extracted using the TRIzol((TM)) method. This was followed by detection using SYBR Green I real-time RT-PCR with primers specific for NoV GI (COG1F-COG1R) and GII (COG2F-COG2R) along with an internal RNA amplification control. End-point detection limits from lettuce and tomatoes were found to be 10 RT-PCR units/25 g for GI and GII and 1 RT-PCR unit/25 g for GI and 10 RT-PCR units/25 g for GII from green onions. These results were confirmed by Tm analysis (showing peaks at 81.5 and 84A degrees C for GI and GII, respectively; and 83A degrees C for the IAC) as well as agarose gel electrophoresis that confirmed products of similar to 95 bp for GI and GII and similar to 155 bp for the RNA IAC. Results could be obtained within one working day, showing potential for routine use in diagnostics and monitoring of NoV contamination by the produce industry.", publisher = "Springer, New York", journal = "Food and Environmental Virology", title = "Simple and Rapid Detection of Human Norovirus from Produce Using SYBR Green I-based Real-time RT-PCR", pages = "129-121", number = "3-4", volume = "3", doi = "10.1007/s12560-011-9066-5" }
Radin, D.,& D'Souza, D. H.. (2011). Simple and Rapid Detection of Human Norovirus from Produce Using SYBR Green I-based Real-time RT-PCR. in Food and Environmental Virology Springer, New York., 3(3-4), 121-129. https://doi.org/10.1007/s12560-011-9066-5
Radin D, D'Souza DH. Simple and Rapid Detection of Human Norovirus from Produce Using SYBR Green I-based Real-time RT-PCR. in Food and Environmental Virology. 2011;3(3-4):121-129. doi:10.1007/s12560-011-9066-5 .
Radin, Dragoslava, D'Souza, Doris H., "Simple and Rapid Detection of Human Norovirus from Produce Using SYBR Green I-based Real-time RT-PCR" in Food and Environmental Virology, 3, no. 3-4 (2011):121-129, https://doi.org/10.1007/s12560-011-9066-5 . .