Comparison of serological and molecular methods for differentiation between genotype A and genotype B strains of small ruminant lentiviruses

Olech, Monika; Kuźmak, Jacek

doi:10.2478/jvetres-2024-0025

Abstract

Introduction

Small ruminant lentiviruses (SRLV) cause multisystemic, degenerative and chronic disease in sheep and goats. There are five genotypes (A, B, C, D and E), of which A and B are the most widespread. The purpose of this study was to evaluate the serotyping efficiency of the Eradikit SRLV Genotyping ELISA and the molecular typing efficiency of a newly developed nested real-time PCR targeting the long terminal repeat–gag (LTR-gag) region using samples from animals infected with subtypes of SRLV known to circulate in Poland.

Material and Methods

A total of 97 sera samples taken from 34 sheep and 63 goats were immunoassayed, and 86 DNA samples from 31 sheep and 55 goats were tested with the PCR. All ruminants were infected with known SRLV strains of the A1, A5, A12, A13, A16, A17, A18, A23, A24, A27, B1 and B2 subtypes.

Results

A total of 69 (80.2%, 95% confidence interval 71.6%–88.8%) out of 86 tested samples gave positive results in the PCR. In 17 out of the 86 (19.8%) samples, no proviral DNA of SRLV was detected. The differentiation between MVV (genotype A) and CAEV (genotype B) by PCR matched the predating phylogenetic analysis invariably. No cross-reactivity was observed. On the other hand, the proportion of samples genotyped the same by the older phylogenetic analysis and the Eradikit SRLV Genotyping ELISA was 42.3%. The test was unable to classify 40.2% of samples, and 17.5% of sera were incorrectly classified.

Conclusion

Our results showed that the Eradikit SRLV genotyping kit is not a reliable method for predicting SRLV genotype, while the nested real-time PCR based on the LTR-gag region did prove to be, at least for genotypes A and B.

References

Acevedo Jiménez G.E., Tórtora Pérez J.L., Rodríguez Murillo C., Arellano Reynoso B., Ramírez Álvarez H.: Serotyping versus genotyping in infected sheep and goats with small ruminant lentiviruses. Vet Microbiol 2021, 252, 10893, doi: 10.1016/j.vetmic.2020.108931.
Open DOI Search in Google Scholar Back to article
Blatti-Cardinaux L., Pisoni G., Stoffel M.H., Zanoni R., Zahno M.L., Bertoni G.: Generation of a molecular clone of an attenuated lentivirus, a first step in understanding cytopathogenicity and virulence. Virology 2016, 487, 50–58, doi: 10.1016/j.virol.2015.09.027.
Open DOI Search in Google Scholar Back to article
Brajon G., Mandas D., Liciardi M., Taccori F., Meloni M., Corrias F., Montaldo C., Coghe F., Casciari C., Giammarioli M., Orrù G.: Development and Field Testing of a Real-Time PCR Assay for Caprine Arthritis-Encephalitis-Virus (CAEV). Open Virol J 2012, 6, 82–90, doi: 10.2174/1874357901206010082.
Open DOI Search in Google Scholar Back to article
Brinkhof J., van Maanen C.: Evaluation of five enzyme-linked immunosorbent assays and an agar gel immunodiffusion test for detection of antibodies to small ruminant lentiviruses. Clin Vaccine Immunol 2007, 14, 1210–1214, doi: 10.1128/CVI.00282-06.
Open DOI Search in Google Scholar Back to article
Brinkhof J.M., van Maanen C., Wigger R., Peterson K., Houwers D.J.: Specific detection of small ruminant lentiviral nucleic acid sequences located in the proviral long terminal repeat and leadergag regions using real-time polymerase chain reaction. J Virol Methods 2008, 147, 338–344, doi: 10.1016/j.jviromet.2007.10.013.
Open DOI Search in Google Scholar Back to article
Cardinaux L., Zahno M.L., Deubelbeiss M., Zanoni R., Vogt H.R., Bertoni G.: Virological and phylogenetic characterization of attenuated small ruminant lentivirus isolates eluding efficient serological detection. Vet Microbiol 2013, 162, 572–581, doi: 10.1016/j.vetmic.2012.11.017.
Open DOI Search in Google Scholar Back to article
Carrozza M.L., Mazzei M., Lacerenza D., Del Chiaro L., Giammarioli M., Marini C., Rutili D., Rosati S., Tolari F.: Seroconversion against SU5 derived synthetic peptides in sheep experimentally infected with different SRLV genotypes. Vet Microbiol 2009, 137, 369–374, doi: 10.1016/j.vetmic.2009.01.032.
Open DOI Search in Google Scholar Back to article
Celer V. Jr., Celer V.: Detection of antibodies to ovine lentivirus using recombinant capsid and transmembrane proteins. J Vet Med B Infect Dis Vet Public Health 2001, 48, 89–95, doi: 10.1046/j.1439-0450.2001.00430.x.
Open DOI Search in Google Scholar Back to article
Chassalevris T., Chaintoutis S.C., Apostolidi E.D., Giadinis N.D., Vlemmas I., Brellou G.D., Dovas C.I.: A highly sensitive seminested real-time PCR utilizing oligospermine-conjugated degenerate primers for the detection of diverse strains of small ruminant lentiviruses. Mol Cell Probes 2020, 51, 101528, doi: 10.1016/j.mcp.2020.101528.
Open DOI Search in Google Scholar Back to article
De Regge N., Cay B.: Development, validation and evaluation of added diagnostic value of a q(RT)-PCR for the detection of genotype A strains of small ruminant lentiviruses. J Virol Methods 2013, 194, 250–257, doi: 10.1016/j.jviromet.2013.09.001.
Open DOI Search in Google Scholar Back to article
Germain K., Valas S.: Distribution and heterogeneity of small ruminant lentivirus envelope subtypes in naturally infected French sheep. Virus Res 2006, 120, 156–162, doi: 10.1016/j.virusres.2006.03.002.
Open DOI Search in Google Scholar Back to article
Gjerset B., Storset A.K., Rimstad E.: Genetic diversity of small-ruminant lentiviruses: characterization of Norwegian isolates of Caprine arthritis encephalitis virus. J Gen Virol 2006, 87, 573–580, doi: 10.1099/vir.0.81201-0.
Open DOI Search in Google Scholar Back to article
Glaria I., Reina R., Ramírez H., de Andrés X., Crespo H., Jauregui P., Salazar E., Luján L., Pérez M.M., Benavides J., Pérez V., Polledo L., García-Marín J.F., Riezu J.I., Borrás F., Amorena B., de Andrés D.: Visna/Maedi virus genetic characterization and serological diagnosis of infection in sheep from a neurological outbreak. Vet Microbiol 2012, 155, 137–146, doi: 10.1016/j.vetmic.2011.08.027.
Open DOI Search in Google Scholar Back to article
Gogolewski R.P., Adams D.S., McGuire T.C., Banks K.L., Cheevers W.P.: Antigenic cross-reactivity between caprine arthritis-encephalitis, visna and progressive pneumonia viruses involves all virion-associated proteins and glycoproteins. J Gen Virol 1985, 66, 1233–1240, doi: 10.1099/0022-1317-66-6-1233.
Open DOI Search in Google Scholar Back to article
Grego E., Bertolotti L., Carrozza M.L., Profiti M., Mazzei M., Tolari F., Rosati S.: Genetic and antigenic characterization of the matrix protein of two genetically distinct ovine lentiviruses. Vet Microbiol 2005, 106, 179–185, doi: 10.1016/j.vetmic.2004.12.007.
Open DOI Search in Google Scholar Back to article
Grego E., Bertolotti L., Quasso A., Profiti M., Lacerenza D., Muz D., Rosati S.: Genetic characterization of small ruminant lentivirus in Italian mixed flocks: evidence for a novel genotype circulating in a local goat population. J Gen Virol 2007, 88, 3423–3427, doi: 10.1099/vir.0.83292-0.
Open DOI Search in Google Scholar Back to article
Grego E., Profiti M., Giammarioli M., Giannino L., Rutili D., Woodall C., Rosati S.: Genetic heterogeneity of small ruminant lentiviruses involves immunodominant epitope of capsid antigen and affects sensitivity of single-strain-based immunoassay. Clin Diagn Lab Immunol 2002, 9, 828–832, doi: 10.1128/cdli.9.4.828-832.2002.
Open DOI Search in Google Scholar Back to article
Herrmann-Hoesing L.M., White S.N., Lewis G.S., Mousel M.R., Knowles D.P.: Development and validation of an ovine progressive pneumonia virus quantitative PCR. Clin Vaccine Immunol 2007, 14, 1274–1278, doi: 10.1128/CVI.00095-07.
Open DOI Search in Google Scholar Back to article
Kaba J., Czopowicz M., Kuzmak J., Olech M., Witkowski L., Moroz-Fik A., Mickiewicz M., Biernacka K., Nalbert T., Bereznowski A., Szalus-Jordanow O., Potârniche A.V., Spinu M., Markowska-Daniel I., Bagnicka E.: A large-scale study on the seroprevalence of small ruminant lentiviral infection in the Polish goat population. Prev Vet Med 2023, 213, 105885, doi: 10.1016/j.prevetmed.2023.105885.
Open DOI Search in Google Scholar Back to article
Kuhar U., Barlic-Maganja D., Grom J.: Development and validation of TaqMan probe based real time PCR assays for the specific detection of genotype A and B small ruminant lentivirus strains. BMC Vet Res 2013, 9, 172, doi: 10.1186/1746-6148-9-172.
Open DOI Search in Google Scholar Back to article
Lacerenza D., Giammarioli M., Grego E., Marini C., Profiti M., Rutili D., Rosati S.: Antibody response in sheep experimentally infected with different small ruminant lentivirus genotypes. Vet Immunol Immunopathol 2006, 112, 264–271, doi: 10.1016/j.vetimm.2006.03.016.
Open DOI Search in Google Scholar Back to article
Michiels R., Van Mael E., Quinet C., Adjadj N.R., Cay A.B., De Regge N.: Comparative Analysis of Different Serological and Molecular Tests for the Detection of Small Ruminant Lentiviruses (SRLVs) in Belgian Sheep and Goats. Viruses 2018, 10, 696, doi: 10.3390/v10120696.
Open DOI Search in Google Scholar Back to article
Minardi da Cruz J.C., Singh D.K., Lamara A., Chebloune Y.: Small ruminant lentiviruses (SRLVs) break the species barrier to acquire new host range. Viruses 2013, 5, 1867–1884, doi: 10.3390/v5071867.
Open DOI Search in Google Scholar Back to article
Minguijón E., Reina R., Pérez M., Polledo L., Villoria M., Ramírez H., Leginagoikoa I., Badiola J.J., García-Marín J.F., de Andrés D., Luján L., Amorena B., Juste R.A.: Small ruminant lentivirus infections and diseases. Vet Microbiol 2015, 181, 75–89, doi: 10.1016/j.vetmic.2015.08.007.
Open DOI Search in Google Scholar Back to article
Mordasini F., Vogt H.R., Zahno M.L., Maeschli A., Nenci C., Zanoni R., Peterhans E., Bertoni G.: Analysis of the antibody response to an immunodominant epitope of the envelope glycoprotein of a lentivirus and its diagnostic potential. J Clin Microbiol 2006, 44, 981–991, doi: 10.1128/JCM.44.3.981-991.2006.
Open DOI Search in Google Scholar Back to article
Nogarol C., Bertolotti L., Klevar S., Profiti M., Gjerset B., Rosati S.: Serological characterization of small ruminant lentivirus: a complete tool for serotyping lentivirus infection in goat. Small Rumin Res 2019, 176, 42–46, doi: 10.1016/j.smallrumres.2019.05.010.
Open DOI Search in Google Scholar Back to article
Olech M.: The genetic variability of small-ruminant lentiviruses and its impact on tropism, the development of diagnostic tests and vaccines and the effectiveness of control programmes. J Vet Res 2023, 67, 479–502, doi: 10.2478/jvetres-2023-0064.
Open DOI Search in Google Scholar Back to article
Olech M., Croisé B., Kuzmak J., Valas S.: Evidence for interspecies transmission of small ruminant lentiviruses in sheep and goats in Poland. Bull Vet Inst Pulawy 2009, 53, 165–168.
Search in Google Scholar Back to article
Olech M., Kuzmak J.: Molecular Characterization of Small Ruminant Lentiviruses of Subtype A5 Detected in Naturally Infected but Clinically Healthy Goats of Carpathian Breed. Pathogens 2020, 9, 992, doi: 10.3390/pathogens9120992.
Open DOI Search in Google Scholar Back to article
Olech M., Kuzmak J.: Molecular Characterization of Small Ruminant Lentiviruses in Polish Mixed Flocks Supports Evidence of Cross Species Transmission, Dual Infection, a Recombination Event, and Reveals the Existence of New Subtypes within Group A. Viruses 2021, 13, 2529, doi: 10.3390/v13122529.
Open DOI Search in Google Scholar Back to article
Olech M., Kycko A., Kuzmak J.: Molecular Characterization of Small Ruminant Lentiviruses Isolated from Polish Goats with Arthritis. Viruses 2022, 14, 735, doi: 10.3390/v14040735.
Open DOI Search in Google Scholar Back to article
Olech M., Murawski M., Kuzmak J.: Molecular analysis of small-ruminant lentiviruses in Polish flocks reveals the existence of a novel subtype in sheep. Arch Virol 2019, 164, 1193–1198, doi: 10.1007/s00705-019-04161-9.
Open DOI Search in Google Scholar Back to article
Olech M., Osński Z., Kuźmak J.: Bayesian estimation of seroprevalence of small ruminant lentiviruses in sheep from Poland. Prev Vet Med 2017, 147, 66–78, doi: 10.1016/j.prevetmed.2017.09.001.
Open DOI Search in Google Scholar Back to article
Olech M., Rachid A., Croisé B., Kuzmak J., Valas S.: Genetic and antigenic characterization of small ruminant lentiviruses circulating in Poland. Virus Res 2012, 163, 528–536, doi: 10.1016/j.virusres.2011.11.019.
Open DOI Search in Google Scholar Back to article
Olech M., Valas S., Kuzmak J.: Epidemiological survey in single-species flocks from Poland reveals expanded genetic and antigenic diversity of small ruminant lentiviruses. PLoS One 2018, 13, e0193892, doi: 10.1371/journal.pone.0193892.
Open DOI Search in Google Scholar Back to article
Reina R., Bertolotti L., Dei Giudici S., Puggioni G., Ponti N., Profiti M., Patta C., Rosati S.: Small ruminant lentivirus genotype E is widespread in Sarda goat. Vet Microbiol 2010, 144, 24–31, doi: 10.1016/j.vetmic.2009.12.020.
Open DOI Search in Google Scholar Back to article
Reina R., Grego E., Profiti M., Glaria I., Robino P., Quasso A., Amorena B., Rosati S.: Development of specific diagnostic test for small ruminant lentivirus genotype E. Vet Microbiol 2009, 138, 251–257, doi: 10.1016/j.vetmic.2009.04.005.
Open DOI Search in Google Scholar Back to article
Rosati S., Mannelli A., Merlo T., Ponti N.: Characterization of the immunodominant cross-reacting epitope of visna maedi virus and caprine arthritis-encephalitis virus capsid antigen. Virus Res 1999, 61, 177–183, doi: 10.1016/s0168-1702(99)00031-3.
Open DOI Search in Google Scholar Back to article
Rosati S., Profiti M., Lorenzetti R., Bandecchi P., Mannelli A., Ortoffi M., Tolari F., Ciabatti I.M.: Development of recombinant capsid antigen/transmembrane epitope fusion proteins for serological diagnosis of animal lentivirus infections. J Virol Methods 2004, 121, 73–78, doi: 10.1016/j.jviromet.2004.06.001.
Open DOI Search in Google Scholar Back to article
Schaer J., Cvetnic Z., Sukalic T., Dörig S., Grisiger M., Iscaro C., Feliziani F., Pfeifer F., Origgi F., Zanoni R.G., Abril C.E.: Evaluation of Serological Methods and a New Real-Time Nested PCR for Small Ruminant Lentiviruses. Pathogens 2022, 11,129, doi: 10.3390/pathogens11020129.
Open DOI Search in Google Scholar Back to article