Interlaboratory Validation of a Method for Hepatitis E Virus RNA Detection in Meat and Meat Products

Althof, Nadine; Trojnar, E.; Böhm, Thomas; Burkhardt, Sabine; Carl, Anja; Contzen, Matthias; Kilwinski, Jochen; Mergemeier, Steffen; Moor, Dominik; Mäde, Dietrich; Johne, Reimar

doi:10.1007/s12560-018-9360-6

Cited by 15 publications

(15 citation statements)

References 34 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Due to the relatively simple cultivation method for this bacteriophage and its usability in laboratories with no restrictions at low biosafety levels, it can easily be propagated in many laboratories. While in some more complex matrices, such as sausages, the use of the bacteriophage MS2 as a process control has been shown to be somewhat problematic [ 15 , 16 , 30 ]; its usability in liver homogenates seems to be possible as determined in our study.…”

Section: Discussionmentioning

confidence: 70%

“…Reliable methods should be suitable for use in routine laboratories, which requires their validation in collaborative studies. So far, only one collaborative study for the validation of a method for HEV-RNA detection in meat products has been published [ 30 ]. However, this method focuses on analysis of pork sausages and is very laborious and time-consuming.…”

Section: Discussionmentioning

confidence: 99%

“…The used method was previously published by Trojnar et al [ 30 ] and can be divided into the three modules sample homogenization, extraction of the RNA and detection of HEV-RNA by real-time RT-PCR ( Figure 1 ). Different amounts of starting material (1 to 10 g) were used for different applications.…”

Section: Methodsmentioning

confidence: 99%

“…Since the samples arrived pre-homogenized, the analysis protocol was started with Step 2 of the homogenization procedure using a sub-sample of 25 mg from each sample ( Figure 1 ). In order to monitor the method performances, several controls were included as described by Althof et al [ 30 ]. Each sample was analyzed in a single reaction, undiluted and in 1:10 dilution.…”

Section: Methodsmentioning

confidence: 99%

“…In line with this, the European Food Safety Authority (EFSA) has recently ranked the development of standardized methods for detection of HEV in meat and meat products as one of the top five priorities of future foodborne virus research [ 29 ]. However, to our knowledge, only one collaborative study for validation of a method for the detection of HEV-RNA in meat products has been published so far, which focused on porcine sausages [ 30 ]. In contrast, no method for HEV-RNA detection in liver samples of pigs and wild boar has been validated for its reproducibility in routine laboratories so far, although this matrix is of high risk for HEV contamination [ 5 , 23 ].…”

Section: Introductionmentioning

confidence: 99%

See 4 more Smart Citations

Interlaboratory Validation of a Detection Method for Hepatitis E Virus RNA in Pig Liver

Trojnar

Contzen

Moor³

et al. 2020

Microorganisms

Self Cite

View full text Add to dashboard Cite

Background: In the last years, the number of notified hepatitis E cases in humans has continuously increased in Europe. Foodborne infection with the zoonotic hepatitis E virus (HEV) genotype 3 is considered the major cause of this disease. Undercooked liver and raw sausages containing the liver of pigs and wild boar are at high risk of containing HEV. However, so far, no standardized method for the detection of HEV-RNA in pig liver is available. Methods: An international collaborative study on method reproducibility involving 11 laboratories was performed for an HEV-RNA detection method, which consists of steps of sample homogenization, RNA extraction and real-time RT-PCR detection, including a process control. Naturally contaminated pork liver samples containing two different amounts of HEV and a HEV-negative pork liver sample were tested by all laboratories using the method. Results: Valid results were retrieved from 10 laboratories. A specificity of 100% and a sensitivity of 79% were calculated for the method. False negative results were only retrieved from the sample containing very low HEV amounts near the detection limit. Conclusions: The results show that the method is highly specific, sufficiently sensitive and robust for use in different laboratories. The method can, therefore, be applied to routine food control as well as in monitoring studies.

show abstract

Section: Discussionmentioning

confidence: 70%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Interlaboratory Validation of a Detection Method for Hepatitis E Virus RNA in Pig Liver

Trojnar

Contzen

Moor³

et al. 2020

Microorganisms

Self Cite

View full text Add to dashboard Cite

show abstract

Hepatitis E virus cross-contamination on the surface of porcine livers after storage in Euro meat containers in a German pig abattoir

Dzierzon

Oswaldi

Merle

et al. 2021

J Consum Prot Food Saf

View full text Add to dashboard Cite

Hepatitis E virus (HEV) is a foodborne zoonotic pathogen and known as the causative agent of hepatitis E in humans. The specific role of porcine liver as a vehicle for human HEV infections has been highlighted in different studies. Nevertheless, gaps of knowledge still exist regarding possible HEV cross-contamination both at consumer and production level. Furthermore, people working in the food production industry, e.g. veterinarians and abattoir employees, are exposed to an increased risk of HEV infection. The aim of the present study was to investigate HEV cross-contamination on the surface of porcine liver in a German abattoir. The sample set included 250 samples of porcine liver parenchyma and the corresponding 250 superficial layer samples of the same livers, which were analyzed for the presence of HEV ribonucleic acid (RNA). Afterwards, the initial status of the tested liver parenchyma was compared with the occurrence of HEV RNA in the corresponding superficial layer. HEV RNA was detectable in 34% (85/250) of superficial layer samples, with 58% (49/85) of the samples originated from initially HEV negative livers. To our knowledge, this is the first study that provides an insight in the potential of HEV cross-contamination at abattoir level in Germany. Furthermore, it could be identified that the joint storage of livers in Euro meat containers has a significant impact on the presence of HEV RNA on the surface of porcine liver.

show abstract

No Evidence of Hepatitis E Virus Infection in Farmed Deer in Germany

2019

View full text Add to dashboard Cite

Interlaboratory Validation of a Method for Hepatitis E Virus RNA Detection in Meat and Meat Products

Cited by 15 publications

References 34 publications

Interlaboratory Validation of a Detection Method for Hepatitis E Virus RNA in Pig Liver

Interlaboratory Validation of a Detection Method for Hepatitis E Virus RNA in Pig Liver

Hepatitis E virus cross-contamination on the surface of porcine livers after storage in Euro meat containers in a German pig abattoir

No Evidence of Hepatitis E Virus Infection in Farmed Deer in Germany

Contact Info

Product

Resources

About