Chigerwe Munashe, Crossley Beate M
Department of Medicine and Epidemiology, University of California, Davis, Davis, CA 95616, USA.
J Vet Diagn Invest. 2013 Jul;25(4):498-501. doi: 10.1177/1040638713490690. Epub 2013 Jun 18.
The objective of the current study was to investigate if the presence of colostral-derived immunoglobulin G (IgG) in blood is an inhibitor of diagnostic polymerase chain reaction (PCR) for detection of Bovine viral diarrhea virus (BVDV). Eleven precolostral and 11 postcolostral blood samples in ethylenediamine tetra-acetic acid (EDTA) anticoagulant as well as serum samples were collected from 11 Holstein bull calves. Calves were fed 3 liters of colostrum once, by oroesophageal tubing. Postcolostral, blood, and serum samples were collected at 48 hr of age. Serum IgG concentrations were determined in the precolostral and postcolostral serum samples using radial immunodiffusion. The blood samples (precolostral and postcolostral) were spiked with BVDV, and 2 diagnostic PCR extraction methods were applied to each sample. The extraction and amplification efficiencies of the 2 PCR methods on the precolostral and postcolostral EDTA blood samples were evaluated. Two of the 11 calves had inadequate passive transfer of colostral immunoglobulins at 48 hr of age based on the serum IgG concentrations. All blood samples from calves were negative for BVDV prior to the spiking with the virus. Evaluation of the 2 different methods among 3 different virus concentrations demonstrated that there was no difference in extraction or amplification efficiency in precolostral and postcolostral samples. The results of this study suggest that bovine IgG is not an inhibitor of PCR used for detection of BVDV in cattle. The methods used in the current study are acceptable for PCR detection of BVDV in cattle.
本研究的目的是调查血液中初乳来源的免疫球蛋白G(IgG)是否会抑制用于检测牛病毒性腹泻病毒(BVDV)的诊断性聚合酶链反应(PCR)。从11头荷斯坦公牛犊中采集了11份采前和11份采后用乙二胺四乙酸(EDTA)抗凝的血液样本以及血清样本。通过食管插管一次性给犊牛饲喂3升初乳。在犊牛48小时龄时采集采后血液和血清样本。使用放射免疫扩散法测定采前和采后血清样本中的血清IgG浓度。将血液样本(采前和采后)接种BVDV,并对每个样本应用2种诊断性PCR提取方法。评估了2种PCR方法对采前和采后EDTA血液样本的提取和扩增效率。根据血清IgG浓度,11头犊牛中有2头在48小时龄时初乳免疫球蛋白的被动转移不足。在接种病毒之前,犊牛的所有血液样本BVDV检测均为阴性。在3种不同病毒浓度下对2种不同方法进行评估,结果表明采前和采后样本的提取或扩增效率没有差异。本研究结果表明,牛IgG不是用于检测牛BVDV的PCR的抑制剂。本研究中使用的方法可用于牛BVDV的PCR检测。