title: Saliva as an alternative specimen for detection of Schmallenberg virus-specific antibodies in bovines.
abstract: BACKGROUND
  Schmallenberg virus (SBV), discovered in continental Europe in late 2011, causes mild clinical signs in adult ruminants, including diarrhoea and reduced milk yield. However, fetal infection can lead to severe malformation in newborn offspring. Enzyme-linked immunosorbent assays (ELISA) are commercially available for detection of SBV-specific antibodies in bovine sera and milk. Here we describe the development and evaluation of an indirect ELISA based on a yeast derived recombinant SBV nucleocapsid protein (N) for the detection of SBV-specific antibodies in bovine saliva. Development of a non-invasive test to detect antibodies in individual bovine saliva samples could potentially provide a test suitable for calves and adult cattle. The aim of this study was to investigate the agreement between the levels of antibodies (IgG) measured in milk and sera, and the level of antibodies (IgG and IgA) in saliva, in comparison with the antibody levels detected in sera and milk with commercially available test.
  
  
  RESULTS
  Serum, milk and saliva samples from 58 cows were collected from three dairy herds in Lithuania and tested for the presence of SBV-specific antibodies. The presence of IgG antibodies was tested in parallel serum and milk samples, while the presence of IgA and IgG antibodies was tested in saliva samples. The presence of SBV-specific IgG and IgA in saliva was tested using an indirect ELISA based on a yeast-derived recombinant N protein. The presence of SBV-specific IgG in milk and sera was tested in parallel using a commercial recombinant protein based test. The sensitivities of the newly developed tests were as follows: 96 % for the IgG serum assay and 94 % for the IgG milk assay and 85 % and 98 % for IgG and IgA in saliva tests, when compared with data generated by a commercial IgG assay.
  
  
  CONCLUSIONS
  Data from testing the saliva IgG and IgA and also the milk and serum IgG with indirect SBV-specific ELISAs showed close agreement with the commercial serum and milk IgG assay data. The level of IgG in saliva was notably lower in comparison to IgA. The newly developed method exhibits the potential to serve as an easily transferable tool for epidemiological studies.
authors.affiliation: Institute of Biotechnology, Vilnius University, V. A. Graiciuno 8, 02241, Vilnius, Lithuania. justas.lazutka@bti.vu.lt.
authors.forename: Justas
authors.initials: J
authors.lastname: Lazutka
authors.affiliation: Institute of Biotechnology, Vilnius University, V. A. Graiciuno 8, 02241, Vilnius, Lithuania. aliona.spakova@gf.stud.vu.lt.
authors.forename: Aliona
authors.initials: A
authors.lastname: Spakova
authors.affiliation: Institute of Microbiology and Virology, Veterinary Academy, Lithuanian University of Health Sciences, Tilzes 18, 47181, Kaunas, Lithuania. sereika@lva.lt.
authors.forename: Vilimas
authors.initials: V
authors.lastname: Sereika
authors.affiliation: Institute of Microbiology and Virology, Veterinary Academy, Lithuanian University of Health Sciences, Tilzes 18, 47181, Kaunas, Lithuania. lelesius@lva.lt.
authors.forename: Raimundas
authors.initials: R
authors.lastname: Lelesius
authors.affiliation: Institute of Biotechnology, Vilnius University, V. A. Graiciuno 8, 02241, Vilnius, Lithuania. kestutis.sasnauskas@bti.vu.lt.
authors.forename: Kestutis
authors.initials: K
authors.lastname: Sasnauskas
authors.affiliation: Institute of Biotechnology, Vilnius University, V. A. Graiciuno 8, 02241, Vilnius, Lithuania. rasa.burneikiene@bti.vu.lt.
authors.forename: Rasa
authors.initials: R
authors.lastname: Petraityte-Burneikiene
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000737:chemistry*; D002044:Bunyaviridae Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis / Q000821:virology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary; D007070:Immunoglobulin A / Q000737:chemistry; D007074:Immunoglobulin G / Q000737:chemistry; D016851:Orthobunyavirus / Q000276:immunology*; D012463:Saliva / Q000737:chemistry*
chemical_list: D000914:Antibodies, Viral; D007070:Immunoglobulin A; D007074:Immunoglobulin G