No Evidence for Xenotropic Murine Leukemia-Related Virus Infection in Sweden Using Internally Controlled Multiepitope Suspension Array Serology

Abstract

<jats:title>ABSTRACT</jats:title><jats:p>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<jats:italic>env</jats:italic>and<jats:italic>gag</jats:italic>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.</jats:p>

Serology in the Digital Age: Using Long Synthetic Peptides Created from Nucleic Acid Sequences as Antigens in Microarrays

“…Antibodies to a pathogen are traditionally detected to one antigen at a time using agent-specific ELISA (enzyme-linked immunosorbent assays). However, many different antibodies can now be detected simultaneously, e.g., with SMIA (suspension multiplex immunoassay) [ 7 , 8 , 9 , 10 ], a rational and economic technique. SMIA is a cost-effective serological method where antigen and serum consumption can be minimized, its background is generally low, and the dynamic range wider than in ELISA.…”

“…SMIA is a cost-effective serological method where antigen and serum consumption can be minimized, its background is generally low, and the dynamic range wider than in ELISA. Unlike the latter, it provides multiple results per analysis [ 7 , 8 , 9 , 10 ].…”

Serological evaluation of possible exposure to Ljungan virus and related parechovirus in autoimmune (type 1) diabetes in children

“…The aim of this study was to compare IgM‐ and IgG‐antibodies to both LV and HPeV peptides detected in an exploratory indirect antibody capture immunoassay [Blomberg et al, ] with LV antibodies detected in a RBA [Niklasson et al, , 2009, 2013] and to test whether these LV antibodies were related to islet autoantibodies or to type 1 diabetes‐associated HLA‐DQ genotypes.…”

No Evidence of Xenotropic Murine Leukemia Virus-Related Virus Transmission by Blood Transfusion from Infected Rhesus Macaques

“…T he initial discovery of xenotropic murine leukemia virus-related virus (XMRV) in human prostate cancer tissue (1), and later in some peripheral blood mononuclear cell (PBMC) samples from patients with myalgic encephalomyelitis/chronic fatigue syndrome (2), raised public health concerns related to the discovery of a novel human retrovirus and potential virus transmission due to exposure to mice (3) and infected blood donors and bloodderived products (4)(5)(6)(7)(8)(9)(10)(11). These emerging concerns led to intensive discussions and investigations of XMRV, including molecular and biological characterization of the virus and the development of assays, standards, and nonhuman primate (NHP) models for further studies of human infection and disease association.…”