Quantification of hepatitis E virus in raw pork livers using droplet digital RT-PCR

Wang, Kairui; Liu, Libing; Wang, Jinfeng; Sun, Xiaoxia; Han, Qian; Zhou, Cang; Xu, Xiangdong; Wang, Jianchang

doi:10.1016/j.fm.2022.104114

Cited by 8 publications

(2 citation statements)

References 35 publications

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“…Even though previous studies applied the ddPCR for the quantification of HEV, none provided a co-amplification of process control [ 17 , 40 , 41 , 42 , 43 ].…”

Section: Discussionmentioning

confidence: 99%

Duplex Droplet Digital PCR Assay for Quantification of Hepatitis E Virus in Food

La Bella,

Basanisi,

Nobili

et al. 2024

Viruses

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Hepatitis E virus (HEV) represents an emerging risk in industrialized countries where the consumption of contaminated food plays a pivotal role. Quantitative real-time RT-PCR (RT-qPCR) is one of the most suitable methods for the detection and quantification of viruses in food. Nevertheless, quantification using RT-qPCR has limitations. Droplet digital PCR (ddPCR) provides the precise quantification of nucleic acids without the need for a standard curve and a reduction in the effect on virus quantification due to the presence of inhibitors. The objectives of the present work were (i) to develop a method for the absolute quantification of HEV in swine tissues based on ddPCR technology and provide internal process control for recovery assessment and (ii) to evaluate the performance of the method by analyzing a selection of naturally contaminated wild boar muscle samples previously tested using RT-qPCR. The method was optimized using a set of in vitro synthesized HEV RNA and quantified dsDNA. The limit of detection of the developed ddPCR assay was 0.34 genome copies/µL. The analysis of the wild boar samples confirmed the validity of the ddPCR assay. The duplex ddPCR method showed no reduction in efficiency compared to individual assays. The method developed in the present study could represent a sensitive assay for the detection and absolute quantification of HEV RNA in food samples with the advantage of presenting the co-amplification of internal process control.

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“…Even though previous studies applied the ddPCR for the quantification of HEV, none provided a co-amplification of process control [ 17 , 40 , 41 , 42 , 43 ].…”

Section: Discussionmentioning

confidence: 99%

Duplex Droplet Digital PCR Assay for Quantification of Hepatitis E Virus in Food

La Bella,

Basanisi,

Nobili

et al. 2024

Viruses

View full text Add to dashboard Cite

show abstract

“…In recent years, digital PCR (dPCR) has been recognised as a useful alternative to qPCR for detecting and quantifying viruses in food samples [ 205 , 206 , 207 ]. The idea of digital PCR was described already in the 1990s (then referred to as limited dilution PCR [ 208 ]), but it was not until the 2010s that the technique became widely commercially available.…”

Section: Methods Used For Detection Of Hav and Hev In Different Food ...mentioning

confidence: 99%

Hepatitis A Virus and Hepatitis E Virus as Food- and Waterborne Pathogens—Transmission Routes and Methods for Detection in Food

Nemes

Persson

Simonsson

2023

Viruses

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Foodborne viruses are an important threat to food safety and public health. Globally, there are approximately 5 million cases of acute viral hepatitis due to hepatitis A virus (HAV) and hepatitis E virus (HEV) every year. HAV is responsible for numerous food-related viral outbreaks worldwide, while HEV is an emerging pathogen with a global health burden. The reported HEV cases in Europe have increased tenfold in the last 20 years due to its zoonotic transmission through the consumption of infected meat or meat products. HEV is considered the most common cause of acute viral hepatitis worldwide currently. This review focuses on the latest findings on the foodborne transmission routes of HAV and HEV and the methods for their detection in different food matrices.

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An international inter-laboratory study to compare digital PCR with ISO standardized qPCR assays for the detection of norovirus GI and GII in oyster tissue

Boxman,

Molin,

Persson

et al. 2024

Food Microbiology

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Quantification of hepatitis E virus in raw pork livers using droplet digital RT-PCR

Cited by 8 publications

References 35 publications

Duplex Droplet Digital PCR Assay for Quantification of Hepatitis E Virus in Food

Duplex Droplet Digital PCR Assay for Quantification of Hepatitis E Virus in Food

Hepatitis A Virus and Hepatitis E Virus as Food- and Waterborne Pathogens—Transmission Routes and Methods for Detection in Food

An international inter-laboratory study to compare digital PCR with ISO standardized qPCR assays for the detection of norovirus GI and GII in oyster tissue

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