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Importantly, PCMV infection of the first genetically modified pig heart into a human may have contributed to the reduced survival of the patient. Sensitive and reliable assays for detection of latent PCMV infection are thus indispensable. Here, we report the development of five peptide\u2010induced rabbit antisera specific for PCMV glycoprotein B (gB) and their validation for detection of PCMV in infected pig fallopian tube (PFT) cells by immunofluorescence and electron microscopy (EM). The anti\u2010gB antibodies were also used for detection by Western blot analysis of PCMV purified from the supernatant of infected PFT cells. Sera of infected versus non\u2010infected pigs have been compared. In parallel, PCMV viral load in blood samples of the animals was quantified by a novel highly sensitive nested\u2010PCR and qPCR assay. A combination of four partly overlapping peptides from the gB C\u2010terminus was used to establish a diagnostic ELISA for PCMV gB specific pig antibodies which is able to differentiate infected from non\u2010infected animals and to quantify maternal antibodies in neonates. The combination of a highly sensitive nested PCR for direct virus detection with a sensitive peptide\u2010based ELISA detecting anti\u2010PCMV gB\u2010antibodies, supplemented by Western blot analysis and\/or immunohistochemistry for virus detection will reliably differentiate pigs with active infection, latently infected pigs, and non\u2010infected pigs. It may significantly improve the virologic safety of xenotransplantation.<\/jats:p>","DOI":"10.1111\/xen.12803","type":"journal-article","created":{"date-parts":[[2023,4,30]],"date-time":"2023-04-30T18:13:55Z","timestamp":1682878435000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":20,"title":["PCR and peptide based PCMV detection in pig \u2013 development and application of a combined testing procedure differentiating newly from latent infected pigs"],"prefix":"10.1111","volume":"30","author":[{"given":"Nicole","family":"Fischer","sequence":"first","affiliation":[{"name":"Division of Haematology Cell and Gene Therapy Paul\u2010Ehrlich\u2010Institut  Langen Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Barbara","family":"Gulich","sequence":"additional","affiliation":[{"name":"Division of Haematology Cell and Gene Therapy Paul\u2010Ehrlich\u2010Institut  Langen 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