Blomberg Jonas, Blomberg Fredrik, Sjösten Anna, Sheikholvaezin Ali, Bölin-Wiener Agnes, Elfaitouri Amal, Hessel Sanna, Gottfries Carl-Gerhard, Zachrisson Olof, Ohrmalm Christina, Jobs Magnus, Pipkorn Rüdiger
Section of Clinical Microbiology, Department of Medical Sciences, Uppsala University, Uppsala, Sweden.
Clin Vaccine Immunol. 2012 Sep;19(9):1399-410. doi: 10.1128/CVI.00391-12. Epub 2012 Jul 11.
Many syndromes have a large number of differential diagnoses, a situation which calls for multiplex diagnostic systems. Myalgic encephalomyelitis (ME), also named chronic fatigue syndrome (CFS), is a common disease of unknown etiology. A mouse retrovirus, xenotropic murine leukemia-related virus (XMRV), was found in ME/CFS patients and blood donors, but this was not corroborated. However, the paucity of serological investigations on XMRV in humans prompted us to develop a serological assay which cover many aspects of XMRV antigenicity. It is a novel suspension array method, using a multiplex IgG assay with nine recombinant proteins from the env and gag genes of XMRV and 38 peptides based on known epitopes of vertebrate gammaretroviruses. IgG antibodies were sought in 520 blood donors and 85 ME/CFS patients and in positive- and negative-control sera from animals. We found no differences in seroreactivity between blood donors and ME/CFS patients for any of the antigens. This did not support an association between ME/CFS and XMRV infection. The multiplex serological system had several advantages: (i) biotinylated protein G allowed us to run both human and animal sera, which is essential because of a lack of XMRV-positive humans; (ii) a novel quality control was a pan-peptide positive-control rabbit serum; and (iii) synthetic XMRV Gag peptides with degenerate positions covering most of the variation of murine leukemia-like viruses did not give higher background than nondegenerate analogs. The principle may be used for creation of variant tolerant peptide serologies. Thus, our system allows rational large-scale serological assays with built-in quality control.
许多综合征都有大量的鉴别诊断，这种情况需要多重诊断系统。肌痛性脑脊髓炎（ME），也称为慢性疲劳综合征（CFS），是一种病因不明的常见疾病。在ME/CFS患者和献血者中发现了一种小鼠逆转录病毒，即嗜异性小鼠白血病相关病毒（XMRV），但这一点并未得到证实。然而，针对人类XMRV的血清学研究较少，这促使我们开发一种涵盖XMRV抗原性多个方面的血清学检测方法。这是一种新型的悬浮阵列方法，使用多重IgG检测，其中包含来自XMRV env和gag基因的九种重组蛋白以及基于脊椎动物γ逆转录病毒已知表位的38种肽。在520名献血者、85名ME/CFS患者以及动物的阳性和阴性对照血清中检测IgG抗体。我们发现，对于任何一种抗原，献血者和ME/CFS患者之间的血清反应性没有差异。这并不支持ME/CFS与XMRV感染之间存在关联。这种多重血清学系统具有几个优点：（i）生物素化的蛋白G使我们能够检测人类和动物血清，这一点至关重要，因为缺乏XMRV阳性的人类样本；（ii）一种新型的质量控制方法是全肽阳性对照兔血清；（iii）具有覆盖大多数鼠白血病样病毒变异的简并位点的合成XMRV Gag肽，其背景并不比非简并类似物更高。该原理可用于创建耐受变异体的肽血清学。因此，我们的系统允许进行具有内置质量控制的合理大规模血清学检测。
