Lewis Joshua, McNab Tegan, Tenaya Masa, Hartaningsih Nining, Wilcox Graham, Desport Moira
School of Veterinary and Biomedical Sciences, Murdoch University, South St., Murdoch, WA 6150, Australia.
J Virol Methods. 2009 Jul;159(1):81-6. doi: 10.1016/j.jviromet.2009.03.005. Epub 2009 Mar 18.
A sensitive diagnostic assay for the detection of infections with the bovine lentivirus Jembrana disease virus (JDV) is required in Indonesia to control the spread of Jembrana disease. Immunoassays are used routinely but are compromised by cross-reactive epitopes in the capsid (CA) protein of JDV and the genetically related bovine immunodeficiency virus (BIV). JDV gag-specific primers were tested in a real-time PCR assay to detect proviral DNA in peripheral blood mononuclear cells from 165 cattle from the Tabanan district of Bali. JDV-specific amplicons were detected in 9% of the cattle and only 33% of the real-time PCR positive cattle were seropositive. The delayed seroconversion that occurs after infection with JDV could explain the low concordance between these assays but other factors may be responsible. BIV proviral DNA was not detected in any of the PBMC DNA samples. A high concordance value of 98.6% was found between the JDV plasma-derived antigen Western blot and the JDV p26-his recombinant protein ELISA. Only 21% of the seropositive cattle had detectable levels of proviral DNA suggesting that the proviral load in recovered cattle is low. A combination of real-time PCR and JDV p26-his ELISA is recommended for the detection of infection with JDV in Indonesia.
在印度尼西亚，为控制杰姆布拉纳病的传播，需要一种灵敏的诊断检测方法来检测牛慢病毒杰姆布拉纳病病毒（JDV）感染。免疫检测方法虽常规使用，但因JDV衣壳（CA）蛋白与基因相关的牛免疫缺陷病毒（BIV）存在交叉反应表位而受到影响。对JDV gag特异性引物进行了实时PCR检测，以检测巴厘岛塔巴南地区165头牛外周血单个核细胞中的前病毒DNA。在9%的牛中检测到了JDV特异性扩增子，且实时PCR阳性的牛中只有33%血清呈阳性。感染JDV后出现的血清转化延迟可能解释了这些检测方法之间的低一致性，但其他因素可能也有影响。在任何外周血单个核细胞DNA样本中均未检测到BIV前病毒DNA。在JDV血浆来源抗原免疫印迹法与JDV p26-组氨酸重组蛋白ELISA之间发现了98.6%的高一致性值。只有21%的血清阳性牛具有可检测水平的前病毒DNA，这表明康复牛中的前病毒载量较低。建议将实时PCR和JDV p26-组氨酸ELISA结合用于印度尼西亚JDV感染的检测。
