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唾液作为检测牛体内施马伦贝格病毒特异性抗体的替代样本。

Saliva as an alternative specimen for detection of Schmallenberg virus-specific antibodies in bovines.

作者信息

Lazutka Justas, Spakova Aliona, Sereika Vilimas, Lelesius Raimundas, Sasnauskas Kestutis, Petraityte-Burneikiene Rasa

机构信息

Institute of Biotechnology, Vilnius University, V. A. Graiciuno 8, 02241, Vilnius, Lithuania.

Institute of Microbiology and Virology, Veterinary Academy, Lithuanian University of Health Sciences, Tilzes 18, 47181, Kaunas, Lithuania.

出版信息

BMC Vet Res. 2015 Sep 15;11:237. doi: 10.1186/s12917-015-0552-0.

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.

摘要

背景

施马伦贝格病毒(SBV)于2011年末在欧洲大陆被发现,可使成年反刍动物出现轻微临床症状,包括腹泻和产奶量下降。然而,胎儿感染可导致新生后代出现严重畸形。酶联免疫吸附测定(ELISA)可用于商业检测牛血清和牛奶中的SBV特异性抗体。在此,我们描述了一种基于酵母衍生重组SBV核衣壳蛋白(N)的间接ELISA的开发与评估,用于检测牛唾液中的SBV特异性抗体。开发一种非侵入性检测方法来检测个体牛唾液样本中的抗体,可能会提供一种适用于犊牛和成年牛的检测方法。本研究的目的是比较用市售检测方法检测血清和牛奶中的抗体水平,与检测唾液中抗体(IgG和IgA)水平,研究牛奶和血清中测得的抗体(IgG)水平与唾液中抗体水平之间的一致性。

结果

从立陶宛的三个奶牛场收集了58头奶牛的血清、牛奶和唾液样本,并检测其中是否存在SBV特异性抗体。在平行的血清和牛奶样本中检测IgG抗体的存在,而在唾液样本中检测IgA和IgG抗体的存在。使用基于酵母衍生重组N蛋白的间接ELISA检测唾液中SBV特异性IgG和IgA的存在。使用基于商业重组蛋白的检测方法平行检测牛奶和血清中SBV特异性IgG的存在。与商业IgG检测产生的数据相比,新开发检测方法的灵敏度如下:IgG血清检测为96%,IgG牛奶检测为94%,唾液检测中IgG和IgA分别为85%和98%。

结论

用间接SBV特异性ELISA检测唾液IgG和IgA以及牛奶和血清IgG的数据,与商业血清和牛奶IgG检测数据显示出密切一致性。与IgA相比,唾液中IgG水平明显较低。新开发的方法具有作为流行病学研究中易于转移的工具的潜力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e935/4570040/5bbd3ce52ea5/12917_2015_552_Fig1_HTML.jpg

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