Comparison of methods for the detection of porcine cytomegalovirus/roseolovirus in relation to biosafety monitoring of xenotransplantation products

Abstract

BackgroundThe porcine cytomegalovirus, a porcine roseolovirus (PCMV/PRV), is widely distributed in pig populations. It has been shown that PCMV/PRV was transmitted by pig xenotransplants to non‐human primates, and significantly reduced the survival time of the recipient. PCMV/PRV was also transmitted during the first transplantation of a pig heart into a human patient. PCMV/PRV establishes a lifelong persistent infection (latency) in the host, is difficult to detect in this stage, and consequential poses a threat to future clinical xenotransplantations. Therefore, sensitive and specific methods and goal‐oriented strategies how, when, and where to test should be used for screening donor pigs.MethodsIn this study we compared experimentally the PCMV/PRV detection methods including PCR‐based (real‐time PCR, nested PCR) and immunological methods (Western blot assay, ELISA) recently published by Halecker et al. (Sci. Rep. 2022;12(1):21545) and Fischer et al. (Xenotransplantation 2023:e12803). We also compared the antigens used for antibody detection (a recombinant protein and synthetic peptides corresponding to a conserved region of the glycoprotein B, gB).ResultsThe published methods can be used for screening donor pigs, with the results being similar. The antigens used for the detection of PCMV/PRV‐specific antibodies are almost identical and give comparable results. Overall, the optimal diagnostic tests, the samples used for testing and the time of sampling play a crucial role in preventing the transmission of PCMV/PRV during xenotransplantation.ConclusionSensitive methods are available to screen donor pigs for PCMV/PRV, but a rational application of a combination of PCR‐based and immunological methods as well as rational detection strategies are important for the detection of the virus during latency.

Screening for Viruses in Indigenous Greek Black Pigs

“…Our assay is specific: negative sera were negative at high s). Furthermore, using this Western blot assay, similar results we with an ELISA using synthetic peptides [35].…”

“…The result is comparable with a Western blot testing of animals from a German slaughterhouse [ 28 ]. The R2 fragment was shown immunodominant in the gB protein [ 28 ] and gave similar results when compared with an ELISA using synthetic peptides corresponding to the R2 sequence [ 35 , 36 ]. Our Western blot assay was used repeatedly to determine the antibody response in different pig breeds [ 29 , 30 ].…”