Detection of porcine endogenous retrovirus (PERV) viremia in diseased versus healthy US pigs by qualitative and quantitative real-time RT-PCR.
ABSTRACT Previous studies have linked levels of porcine endogenous retroviruses (PERV) with poor health and disease in pigs. To determine the levels of expression of PERVs and their potential association with disease expression, real-time reverse transcriptase (RT) PCR assays were used to assess PERV-ABC, PERV-C and PERV-A/C levels in three commercial swine operations in the United States. Pigs (n = 204) aged 3-25 weeks were screened, and all 369 serum samples collected were found to be positive for PERV-ABC RNA as expected. PERV-C and PERV-A/C RNA were detected in 24.1% (89/369) and 18.7% (69/369) of the samples, respectively. When divided into age groups, PERV-A/C RNA was identified in 20.0% (43/215) of the nursery pig samples (3-9 weeks of age) compared to 16.9% (26/154) finisher pig samples (12-25 weeks of age). On two of the farms, serum was collected from healthy pigs (n = 60) and from pen-mates with various clinical conditions including diarrhoea, wasting and respiratory disease (n = 60). Overall, 25% (15/60) of the samples from clinically affected pigs were found to be positive for PERV-A/C RNA, whereas in clinically healthy pigs, only 8.3% (5/60) of the samples were found to be PERV-A/C positive (P = 0.026). It was possible to identify PERV-A/C in the same pigs on more than one consecutive bleeding, indicating variable length of PERV-A/C viremia. The results indicate that there is an increased incidence of PERV-A/C viremia in diseased pigs and that PERV-A/C can be detected over time in selected pigs within commercial pig production systems in the United States.
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ABSTRACT: In the context of the shortage of organs and other tissues for use in human transplantation, xenotransplantation procedures with material taken from pigs have come under increased consideration. However, there are unclear consequences of the potential transmission of porcine pathogens to humans. Of particular concern are porcine endogenous retroviruses (PERVs). Three subtypes of PERV have been identified, of which PERV-A and PERV-B have the ability to infect human cells in vitro. The PERV-C subtype does not show this ability but recombinant PERV-A/C forms have demonstrated infectivity in human cells. In view of the risk presented by these observations, the International Xenotransplantation Association recently indicated the existence of four strategies to prevent transmission of PERVs. This article focuses on the molecular aspects of PERV infection in xenotransplantation and reviews the techniques available for the detection of PERV DNA, RNA, reverse transcriptase activity and proteins, and anti-PERV antibodies to enable carrying out these recommendations. These methods could be used to evaluate the risk of PERV transmission in human recipients, enhance the effectiveness and reliability of monitoring procedures, and stimulate discussion on the development of improved, more sensitive methods for the detection of PERVs in the future.Viruses 05/2014; 6(5):2062-83. DOI:10.3390/v6052062 · 3.28 Impact Factor
Xenotransplantation 04/2013; 20:135-137. DOI:10.1111/xen.12039 · 1.78 Impact Factor
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ABSTRACT: Xenotransplantation can provide a virtually limitless supply of cells, tissues and organs for a variety of therapeutic procedures. Cells and tissues for use in human transplantation procedures could be supplied using material taken from pigs. However, there is a potential risk of transmission of porcine infectious agents, including porcine endogenous retroviruses (PERVs), to a novel human host, with as yet unknown consequences. Three subtypes of PERV have been identified, of which both PERV-A and PERV-B have the ability to infect human cells in vitro. The third subtype, PERV-C, does not show this ability. Recombinant PERV-A/C forms have demonstrated infectivity in human cell culture. Monitoring in xenotransplantation should comprise screening of the source pig herd (PERV-A and PERV-B level expression assessment, PERV-C detection) and screening of recipients (differentiation between PERV transmission and chimerism). The detection of PERVs includes analyses of both DNA and RNA (PCR and RT-PCR), quantitative determination of the level of PERV nucleic acids (real-time PCR and real-time RT-PCR), assessment of reverse transcriptase (RT) activity (RT assays) and viral and recipient protein detection (immunological methods). In summary, all available methods should be used in monitoring of PERVs in xenotransplantation, and caution should be exercised at all stages of monitoring. Such monitoring has enormous significance for eliminating the possibility of transmission of PERV infection, thus contributing to higher levels of safety in xenotransplantation.Reproductive biology 03/2014; 14(1):68-73. DOI:10.1016/j.repbio.2014.01.006 · 1.05 Impact Factor