two PCR assays and one antibody assay
no evidence of XMRV in 204 controls and 230 autism samples (434 samples in total).
25 autistic children born to mothers with CFS (banked samples & new I think)
20 mixed controls including family members of the children assayed, people with fibromyalgia and people with chronic Lyme disease.
Using a real-time PCR assay, we screened an additional 48 South Carolina autism disorder samples, 96 Italian ASD)samples, 61 South Carolina ASD samples and 184 healthy controls.
A novel real-time PCR assay targeting the pol gene in murine leukemia virus (MuLV) and XMRV was used to detect XMRV
The primers and probes for the assay were designed against regions of XMRV that were 100% conserved in the seven sequenced XMRV strains (available in GenBank; accession numbers GQ497344.1, GQ497343.1, NC_007815.1, EF185282.1, DQ399707.1, DQ241302.1, DQ241301.1).
Sensitivity of the assay was determined by running dilutions (600, 60 and 6 copies per reaction) of synthetic DNA of the VP62 XMRV strain in a background of 2.5 μg of genomic DNA. DNA from XMRV-infected 22Rv1 cells was used as an additional validation of the sensitivity of each assay
The 25 ASD samples from children born to mothers with CFS and the 20 mixed control samples were also submitted to the Centers for Disease Control (CDC) for blinded assaying using an XMRV western blot and a nested PCR assay (external primers XPOLOF and XPOLOR; internal primers XPOLIF and XPOLIR as shown in the Switzer, et al publication) that detects both MuLV and XMRV pol gene sequences .