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用于检测多种家养禽类中禽C型副粘病毒特异性抗体的阻断酶联免疫吸附测定法的建立与评估

Development and evaluation of a blocking enzyme-linked immunosorbent assay for detection of avian metapneumovirus type C-specific antibodies in multiple domestic avian species.

作者信息

Turpin Elizabeth A, Lauer Dale C, Swayne David E

机构信息

Southeast Poultry Research Laboratory, Agricultural Research Service, US Department of Agriculture, Athens, Georgia 30605, USA.

出版信息

J Clin Microbiol. 2003 Aug;41(8):3579-83. doi: 10.1128/JCM.41.8.3579-3583.2003.

Abstract

The first cases of infection caused by avian metapneumoviruses (aMPVs) were described in turkeys with respiratory disease in South Africa during 1978. The causative agent was isolated and identified as a pneumovirus in 1986. aMPVs have been detected in domestic nonpoultry species in Europe, but tests for the detection of these viruses are not available in the United States. To begin to understand the potential role of domestic ducks and geese and wild waterfowl in the epidemiology of aMPV, we have developed and evaluated a blocking enzyme-linked immunosorbent assay (bELISA) for the detection of aMPV type C (aMPV-C)-specific antibodies. This assay method overcomes the species-specific platform of indirect ELISAs to allow detection of aMPV-C-specific antibodies from potentially any avian species. The bELISA was initially tested with experimental turkey serum samples, and the results were found to correlate with those of virus neutralization assays and indirect enzyme-linked immunosorbent assay (iELISA). One thousand serum samples from turkey flocks in Minnesota were evaluated by our bELISA, and the level of agreement of the results of the bELISA and those of the iELISA was 94.9%. In addition, we were able to show that the bELISA could detect aMPV-C-specific antibodies from experimentally infected ducks, indicating its usefulness for the screening of serum samples from multiple avian species. This is the first diagnostic assay for the detection of aMPV-C-specific antibodies from multiple avian species in the United States.

摘要

1978年,南非首次报道了由禽偏肺病毒(aMPV)引起的感染病例,患病火鸡出现呼吸道疾病。1986年,病原体被分离出来并鉴定为一种肺病毒。在欧洲,已经在家养非家禽物种中检测到aMPV,但美国尚无检测这些病毒的试验。为了初步了解家鸭、家鹅和野生水禽在aMPV流行病学中的潜在作用,我们开发并评估了一种阻断酶联免疫吸附测定法(bELISA),用于检测C型aMPV(aMPV-C)特异性抗体。这种测定方法克服了间接ELISA的物种特异性平台,从而能够检测来自任何禽类的aMPV-C特异性抗体。bELISA最初用实验性火鸡血清样本进行检测,结果发现与病毒中和试验和间接酶联免疫吸附测定法(iELISA)的结果相关。我们用bELISA对明尼苏达州火鸡群的1000份血清样本进行了评估,bELISA结果与iELISA结果的一致性水平为94.9%。此外,我们能够证明bELISA可以检测出实验感染鸭的aMPV-C特异性抗体,这表明它可用于筛查多种禽类的血清样本。这是美国首个用于检测多种禽类aMPV-C特异性抗体的诊断测定法。

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