scholarly journals Norovirus Extraction from Frozen Raspberries Using Magnetic Silica Beads

2021 ◽  
Author(s):  
Philippe Raymond ◽  
Sylvianne Paul ◽  
André Perron ◽  
Louise Deschênes
Keyword(s):  
Reverse Transcriptase ◽  
Real Time ◽  
Acute Gastroenteritis ◽  
New Approach ◽  
Human Noroviruses ◽  
Reverse Transcriptase Pcr ◽  
Silica Bead ◽  
Silica Beads ◽  
Magnetic Silica

AbstractHuman noroviruses (HuNoV) are among the main causes of acute gastroenteritis worldwide. Frozen raspberries have been linked to several HuNoV food-related outbreaks. However, the extraction of HuNoV RNA from frozen raspberries remains challenging. Recovery yields are low, and real-time quantitative reverse transcriptase PCR (RT-qPCR) inhibitors limit the sensitivity of the detection methodologies. A new approach using fine magnetic silica beads was developed for the extraction of HuNoV spiked on frozen raspberries. Relatively low recovery yields were observed with both the magnetic silica bead and the reference ISO 15216-1:2017 methods. High RT-qPCR inhibition was observed with the ISO 15216-1:2017 recommended amplification kit but could be reduced by using an alternative kit. Reducing RT-qPCR inhibition is important to limit the number of inconclusive HuNoV assays thus increasing the capacity to assess the HuNoV prevalence in frozen raspberries.

Validation of a Real-Time Reverse Transcriptase–PCR Assay for the Detection of H7 Avian Influenza Virus

2010 ◽  
Vol 5 (s1) ◽  
pp. e148-e149
Author(s):  
Janice Pedersen ◽  
Mary Lea Killian ◽  
Nichole Hines ◽  
Dennis Senne ◽  
Brundaban Panigrahy ◽  
...  
Keyword(s):  
Influenza Virus ◽  
Avian Influenza ◽  
Reverse Transcriptase ◽  
Avian Influenza Virus ◽  
Real Time ◽  
Pcr Assay ◽  
Reverse Transcriptase Pcr

Development of asigDE-Based Real-Time Reverse-Transcriptase PCR for the Detection of ViableSalmonella enterica

2014 ◽  
Vol 11 (7) ◽  
pp. 537-544 ◽  
Author(s):  
Min Zhou ◽  
Jielin Yang ◽  
Xiujuan Zhou ◽  
Bin Liu ◽  
Daixin Liu ◽  
...  
Keyword(s):  
Reverse Transcriptase ◽  
Real Time ◽  
Reverse Transcriptase Pcr

scholarly journals Comparison of Rapid Antigen Test and Real-Time Reverse Transcriptase PCR for Diagnosing Novel Swine Influenza A (H1N1)

2010 ◽  
Vol 13 (3) ◽  
pp. 109 ◽  
Author(s):  
Aerin Kwon ◽  
Jae-Seok Kim ◽  
Han-Sung Kim ◽  
Wonkeun Song ◽  
Ji-Young Park ◽  
...  
Keyword(s):  
Reverse Transcriptase ◽  
Real Time ◽  
Influenza A ◽  
Swine Influenza ◽  
Antigen Test ◽  
Reverse Transcriptase Pcr ◽  
Influenza A H1n1

Novel Approach for Detecting Prohibited Species-Specific Central Nervous System Tissue Contamination in Meat by One-Step Real-Time Reverse Transcriptase PCR

2009 ◽  
Vol 72 (5) ◽  
pp. 1063-1069 ◽  
Author(s):  
M. M. NAGARAJAN ◽  
D. LONGTIN ◽  
C. SIMARD
Keyword(s):  
Central Nervous System ◽  
Nervous System ◽  
Reverse Transcriptase ◽  
Real Time ◽  
Compliance Monitoring ◽  
Reverse Transcriptase Pcr ◽  
Qrt Pcr ◽  
Tissue Contamination ◽  
One Step ◽  
Species Specific

The dissemination of prohibited species-specific central nervous system (CNS) tissue contamination in meat must be tracked to mitigate human health risk associated with bovine spongiform encephalopathy. The efficiency of compliance monitoring and risk control measures taken by concerned regulatory authorities at meat production facilities to avoid such contamination depends on the ability to detect CNS tissue with a reliable and adequately sensitive quantitative method. A rapid and convenient one-step real-time quantitative reverse transcriptase PCR (qRT-PCR) assay was developed based on the absolute quantification of glial fibrillary acidic protein (GFAP) mRNA as a marker for CNS tissue contamination in meat. The GFAP RNA quantity corresponding to a percentage of CNS tissue in artificially spiked meat was determined using an appropriate in vitro transcribed target GFAP RNA as a calibration standard in the assay. The assay had a linear dynamic range of 102 to 109 copies of target RNA and was able to detect 0.01% CNS contamination in meat. Further evaluation consisted of an analysis of 272 random meat cuts from carcasses and 109 ground meat samples received from a federally inspected abattoir and two meat processing facilities, respectively, over a 5-month period. The analyzed samples were all negative for CNS tissue contamination at an arbitrarily set lower threshold of 0.025%. Overall, the newly developed one-step qRT-PCR may be useful as an objective quantitative compliance monitoring tool and for setting an acceptable low tolerance threshold for such contamination in meat.

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