Reliable Polymerase Chain Reaction Methods for Screening for Porcine Endogenous Retroviruses-C (PERV-C) in Pigs
Porcine endogenous retrovirus C (PERV-C) is a gammaretrovirus present in the genome of many, but not all, pigs. It is an ecotropic virus, able to infect only pig cells. In contrast, PERV-A and PERV-B, which are present in all pigs, can infect cells of multiple host species, including humans, thereby...
Saved in:
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2025-01-01
|
| Series: | Viruses |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1999-4915/17/2/164 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1850229842816008192 |
|---|---|
| author | Hina Jhelum Dusan Kunec Vasileios Papatsiros Benedikt B. Kaufer Joachim Denner |
| author_facet | Hina Jhelum Dusan Kunec Vasileios Papatsiros Benedikt B. Kaufer Joachim Denner |
| author_sort | Hina Jhelum |
| collection | DOAJ |
| description | Porcine endogenous retrovirus C (PERV-C) is a gammaretrovirus present in the genome of many, but not all, pigs. It is an ecotropic virus, able to infect only pig cells. In contrast, PERV-A and PERV-B, which are present in all pigs, can infect cells of multiple host species, including humans, thereby posing a risk for xenotransplantation when pigs are used as donor animals. Notably, PERV-C can recombine with PERV-A to produce PERV-A/C recombinants that can infect human cells and replicate to higher titers compared to the paternal PERV-A. The objective of this study is to evaluate the reliability of both existing and newly developed polymerase chain reactions (PCR) methods for detecting PERV-C, with the aim of selecting PERV-C-free pigs to be used for xenotransplantation. To detect PERV-C by PCR, specific primers targeting the region of the envelope protein gene, which differs from that of PERV-A and PERV-B due to its unique receptor binding site, must be employed. In this study, new PCR assays were developed to detect PERV-C and a total of ten PCR assays and one real-time PCR assay were evaluated for their reliability in detecting PERV-C. These assays were used to screen indigenous Greek black pigs, Auckland Island pigs, and German slaughterhouse pigs. Two of the PCR assays consistently yielded reliable results, whereas the other PCRs and the real-time PCR gave false positive results. Using the reliable assays, it was shown that one out of four indigenous Greek black pigs (using the same method in a previous publication 11 of 21 pigs were found PERV-C-negative), one out of ten German slaughterhouse pigs, the pig kidney cell line PK15, and all the Auckland Island pigs were PERV-C-negative. The reliable PCR assays will enable the screening of PERV-C-negative donor pigs to be used in xenotransplantation. Most importantly, all the Auckland Island pigs that were genetically modified in Germany for use in clinical trials were PERV-C-negative. |
| format | Article |
| id | doaj-art-d136d29ed6a34f2db45fbd5d47266174 |
| institution | OA Journals |
| issn | 1999-4915 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj-art-d136d29ed6a34f2db45fbd5d472661742025-08-20T02:04:05ZengMDPI AGViruses1999-49152025-01-0117216410.3390/v17020164Reliable Polymerase Chain Reaction Methods for Screening for Porcine Endogenous Retroviruses-C (PERV-C) in PigsHina Jhelum0Dusan Kunec1Vasileios Papatsiros2Benedikt B. Kaufer3Joachim Denner4Institute of Virology, Free University Berlin, 14163 Berlin, GermanyInstitute of Virology, Free University Berlin, 14163 Berlin, GermanyFaculty of Veterinary Medicine, Clinic of Medicine (Farm Animal Medicine), University of Thessaly, GR 43100 Karditsa, GreeceInstitute of Virology, Free University Berlin, 14163 Berlin, GermanyInstitute of Virology, Free University Berlin, 14163 Berlin, GermanyPorcine endogenous retrovirus C (PERV-C) is a gammaretrovirus present in the genome of many, but not all, pigs. It is an ecotropic virus, able to infect only pig cells. In contrast, PERV-A and PERV-B, which are present in all pigs, can infect cells of multiple host species, including humans, thereby posing a risk for xenotransplantation when pigs are used as donor animals. Notably, PERV-C can recombine with PERV-A to produce PERV-A/C recombinants that can infect human cells and replicate to higher titers compared to the paternal PERV-A. The objective of this study is to evaluate the reliability of both existing and newly developed polymerase chain reactions (PCR) methods for detecting PERV-C, with the aim of selecting PERV-C-free pigs to be used for xenotransplantation. To detect PERV-C by PCR, specific primers targeting the region of the envelope protein gene, which differs from that of PERV-A and PERV-B due to its unique receptor binding site, must be employed. In this study, new PCR assays were developed to detect PERV-C and a total of ten PCR assays and one real-time PCR assay were evaluated for their reliability in detecting PERV-C. These assays were used to screen indigenous Greek black pigs, Auckland Island pigs, and German slaughterhouse pigs. Two of the PCR assays consistently yielded reliable results, whereas the other PCRs and the real-time PCR gave false positive results. Using the reliable assays, it was shown that one out of four indigenous Greek black pigs (using the same method in a previous publication 11 of 21 pigs were found PERV-C-negative), one out of ten German slaughterhouse pigs, the pig kidney cell line PK15, and all the Auckland Island pigs were PERV-C-negative. The reliable PCR assays will enable the screening of PERV-C-negative donor pigs to be used in xenotransplantation. Most importantly, all the Auckland Island pigs that were genetically modified in Germany for use in clinical trials were PERV-C-negative.https://www.mdpi.com/1999-4915/17/2/164porcine endogenous retroviruses (PERVs)PERV-Cpolymerase chain reactions (PCR) methodsreal-time PCRxenotransplantationporcine viruses |
| spellingShingle | Hina Jhelum Dusan Kunec Vasileios Papatsiros Benedikt B. Kaufer Joachim Denner Reliable Polymerase Chain Reaction Methods for Screening for Porcine Endogenous Retroviruses-C (PERV-C) in Pigs Viruses porcine endogenous retroviruses (PERVs) PERV-C polymerase chain reactions (PCR) methods real-time PCR xenotransplantation porcine viruses |
| title | Reliable Polymerase Chain Reaction Methods for Screening for Porcine Endogenous Retroviruses-C (PERV-C) in Pigs |
| title_full | Reliable Polymerase Chain Reaction Methods for Screening for Porcine Endogenous Retroviruses-C (PERV-C) in Pigs |
| title_fullStr | Reliable Polymerase Chain Reaction Methods for Screening for Porcine Endogenous Retroviruses-C (PERV-C) in Pigs |
| title_full_unstemmed | Reliable Polymerase Chain Reaction Methods for Screening for Porcine Endogenous Retroviruses-C (PERV-C) in Pigs |
| title_short | Reliable Polymerase Chain Reaction Methods for Screening for Porcine Endogenous Retroviruses-C (PERV-C) in Pigs |
| title_sort | reliable polymerase chain reaction methods for screening for porcine endogenous retroviruses c perv c in pigs |
| topic | porcine endogenous retroviruses (PERVs) PERV-C polymerase chain reactions (PCR) methods real-time PCR xenotransplantation porcine viruses |
| url | https://www.mdpi.com/1999-4915/17/2/164 |
| work_keys_str_mv | AT hinajhelum reliablepolymerasechainreactionmethodsforscreeningforporcineendogenousretrovirusescpervcinpigs AT dusankunec reliablepolymerasechainreactionmethodsforscreeningforporcineendogenousretrovirusescpervcinpigs AT vasileiospapatsiros reliablepolymerasechainreactionmethodsforscreeningforporcineendogenousretrovirusescpervcinpigs AT benediktbkaufer reliablepolymerasechainreactionmethodsforscreeningforporcineendogenousretrovirusescpervcinpigs AT joachimdenner reliablepolymerasechainreactionmethodsforscreeningforporcineendogenousretrovirusescpervcinpigs |