Sequence Analysis of Proviral DNA of Porcine Endogenous Retroviruses

Machnik, Grzegorz; Sypniewski, Daniel; Wydmuch, Z; Cholewa, Krzysztof; Mazurek, Urszula; Wilczok, Tadeusz; Smorąg, Z.; Pacha, J.

doi:10.1016/j.transproceed.2005.10.115

Cited by 5 publications

(7 citation statements)

References 18 publications

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“…The regulatory signals required for retroviral transcription, replication and integration into the host cellular DNA is located within the PERV long terminal repeat (LTR) sequence. Sequence analysis showed that most PERV proviral DNA was signifi cantly mutated, thus suggesting the inability to express functional viral RNA [11].…”

Section: Discussionmentioning

confidence: 99%

Polymerase chain reaction in detection of porcine endogenous retrovirus (PERV) from porcine tissues

Prabha

Verghese

2009

Indian J Microbiol

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Pigs offer an unlimited source of xenografts for humans. The use of transplants from animal origin offers a potential solution to the limited supply of human organs and tissues. However, like many other mammalian species, pigs harbor porcine endogenous retrovirus (PERV), which are encoded in their genomic DNA and are assumed to have been integrated into the porcine germline. The ability of PERV to infect human cells in vitro has heightened safety concerns regarding the transmission of PERV to pig xenograft recipients. Porcine tissues were analyzed using validated assays specifi c for PERV: polymerase chain reaction (PCR) (for PERV DNA) and reverse transcriptase (RT)-PCR (for PERV RNA). PERV-specifi c gag sequences were found in the porcine heart tissue samples using DNA-PCR and RT-PCR. PCR is a rapid and specifi c test for the detection of PERV from xenografts. These fi ndings have demonstrated that the presence of both DNA and RNA forms of PERV in porcine tissues needs to be carefully considered when the infectious disease potential of xenotransplantation is being assessed.

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Section: Discussionmentioning

confidence: 99%

Polymerase chain reaction in detection of porcine endogenous retrovirus (PERV) from porcine tissues

Prabha

Verghese

2009

Indian J Microbiol

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“…This allows characterization of full-length proviral DNA, comparison with sequences obtained for other PERVs, and evaluation of the potential of particular PERVs to undergo recombination events [47,65,91,92]. Sypniewski et al [47] and Machnik et al [65] used long-range PCR to detect full-length PERV DNA and to discriminate them from defective sequences, PCR primer selection depends on the question to be answered. Only when PCR is carried out with the simultaneous use of primers from all of the groups is it possible to assess the ability of the virus to express functional viral RNA [65].…”

Section: The Four Strategies To Prevent Transmission Of Pervsmentioning

confidence: 99%

“…Sypniewski et al [47] and Machnik et al [65] used long-range PCR to detect full-length PERV DNA and to discriminate them from defective sequences, PCR primer selection depends on the question to be answered. Only when PCR is carried out with the simultaneous use of primers from all of the groups is it possible to assess the ability of the virus to express functional viral RNA [65]. This possibility is afforded by multiplex PCR.…”

Section: The Four Strategies To Prevent Transmission Of Pervsmentioning

confidence: 99%

“…Machnik et al [65] showed that most PERV proviral DNA in pig blood samples was significantly mutated. It was also determined that some animals in pig herds can be PERV transmitters or non-transmitters, based on their transmission to human cells in vitro [21,59,61,95].…”

Section: The Four Strategies To Prevent Transmission Of Pervsmentioning

confidence: 99%

“…Some endogenous retroviruses can induce diseases, but they are generally nonpathogenic in their original hosts. Moreover, many endogenous proviral elements are transcriptionally silent or defective, carrying deletions or point mutations, and are thus incapable of producing an infectious virus [64,65]. However, some gammaretroviridae, such as feline leukemia virus, murine leukemia virus, gibbon ape leukemia virus, and koala retrovirus induce leukemia and immunodeficiency in the infected host [26].…”

Section: Introductionmentioning

confidence: 99%

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Porcine Endogenous Retroviruses in Xenotransplantation—Molecular Aspects

Kimsa

Strzałka‐Mrozik

Kimsa

et al. 2014

Viruses

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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.

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Quantitative Analysis of Porcine Endogenous Retroviruses in Different Organs of Transgenic Pigs Generated for Xenotransplantation

et al. 2013

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The pig appears to be the most promising animal donor of organs for use in human recipients. Among several types of pathogens found in pigs, one of the greatest problems is presented by porcine endogenous retroviruses (PERVs). Screening of the source pig herd for PERVs should include analysis of both PERV DNA and RNA. Therefore, the present study focuses on quantitative analysis of PERVs in different organs such as the skin, heart, muscle, and liver and blood of transgenic pigs generated for xenotransplantation. Transgenic pigs were developed to express the human α-galactosidase, the human α-1,2-fucosyltransferase gene, or both genetic modifications of the genome (Lipinski et al., Medycyna Wet 66:316-322, 2010; Lipinski et al., Ann Anim Sci 12:349-356, 2012; Wieczorek et al., Medycyna Wet 67:462-466, 2011). The copy numbers of PERV DNA and RNA were evaluated using real-time Q-PCR and QRT-PCR, respectively. Comparative analysis of all PERV subtypes revealed the following relationships: PERV A > PERV B > PERV C. PERV A and B were found in all samples, whereas PERV C was detected in 47 % of the tested animals. The lowest level of PERV DNA was shown in the muscles for PERV A and B and in blood samples for PERV C. The lowest level of PERV A RNA was found in the skin, whereas those of PERV B and C RNA were found in liver specimens. Quantitative analysis revealed differences in the copy number of PERV subtypes between various organs of transgenic pigs generated for xenotransplantation. Our data support the idea that careful pig selection for organ donation with low PERV copy number may limit the risk of retrovirus transmission to the human recipients.

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Sequence Analysis of Proviral DNA of Porcine Endogenous Retroviruses

Cited by 5 publications

References 18 publications

Polymerase chain reaction in detection of porcine endogenous retrovirus (PERV) from porcine tissues

Polymerase chain reaction in detection of porcine endogenous retrovirus (PERV) from porcine tissues

Porcine Endogenous Retroviruses in Xenotransplantation—Molecular Aspects

Quantitative Analysis of Porcine Endogenous Retroviruses in Different Organs of Transgenic Pigs Generated for Xenotransplantation

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