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一种用于检测猪血清中猪繁殖与呼吸综合征病毒抗体的间接荧光抗体试验。

An indirect fluorescent antibody test for the detection of antibody to swine infertility and respiratory syndrome virus in swine sera.

作者信息

Yoon I J, Joo H S, Christianson W T, Kim H S, Collins J E, Morrison R B, Dial G D

机构信息

Department of Clinical and Population Sciences, College of Veterinary Medicine, University of Minnesota, St. Paul 55108.

出版信息

J Vet Diagn Invest. 1992 Apr;4(2):144-7. doi: 10.1177/104063879200400205.

Abstract

An indirect fluorescent antibody (IFA) test was developed and standardized to detect and quantitate antibody for swine infertility and respiratory syndrome (SIRS) virus in swine sera. Test results were evaluated using sera of pigs infected both experimentally and naturally with SIRS virus. The IFA test used swine alveolar macrophage (SAM) monolayers prepared in 96-well microplates and infected with SIRS virus. The monolayers were incubated with test sera, washed, and stained with fluorescein isothiocyanate-labeled rabbit anti-swine IgG. After another wash step, the monolayers were examined under a fluorescent microscope. A noninfected SAM control well was included for each sample. The antibody titers for each serum sample were recorded as the highest serum dilutions with specific cytoplasmic fluorescence but no fluorescence in the control wells. To evaluate the test, sera of 4 6-week-old pigs that had been infected with SIRS virus, 2 contact pigs, and 13 experimentally infected sows were used. In the experimentally infected pigs, antibody was first detected at 7 days postexposure (PE) and peaked (1:256-1,024) between 11 and 21 days PE. All 13 sow sera were negative at time of infection but were positive (1:64- greater than or equal to 1:1,024) at 14-26 days PE. Seven hundred twenty sera collected from 25 different swine farms with or without a history of SIRS were also tested. Of 344 sera from 15 swine farms with a clinical history of SIRS, 257 (74.7%) sera had IFA titers greater than or equal to 1:4, whereas 371 (98.7%) of 376 sera from herds with no history of SIRS were negative.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

开发并标准化了一种间接荧光抗体(IFA)检测方法,用于检测和定量猪血清中猪不育与呼吸综合征(SIRS)病毒的抗体。使用经实验感染和自然感染SIRS病毒的猪血清对检测结果进行评估。IFA检测采用在96孔微孔板中制备并感染SIRS病毒的猪肺泡巨噬细胞(SAM)单层。将单层与检测血清一起孵育,洗涤后,用异硫氰酸荧光素标记的兔抗猪IgG染色。再经过一次洗涤步骤后,在荧光显微镜下检查单层。每个样品均设有未感染的SAM对照孔。每个血清样品的抗体效价记录为具有特异性细胞质荧光但对照孔无荧光的最高血清稀释度。为评估该检测方法,使用了4头6周龄感染SIRS病毒的猪、2头接触猪以及13头经实验感染的母猪的血清。在经实验感染的猪中,暴露后7天首次检测到抗体,在暴露后11至21天达到峰值(1:256 - 1:1024)。所有13头母猪的血清在感染时均为阴性,但在暴露后14至26天呈阳性(1:64 - 大于或等于1:1024)。还检测了从25个有或无SIRS病史的不同猪场收集的720份血清。在有SIRS临床病史的15个猪场的344份血清中,257份(74.7%)血清的IFA效价大于或等于1:4,而在无SIRS病史的猪群的376份血清中,371份(98.7%)为阴性。(摘要截短于250字)

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