Published online October 2, 2006
PEDIATRICS Vol. 118 No. 4 October 2006, pp. 1664-1675 (doi:10.1542/peds.2006-1262)
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ARTICLE

No Evidence of Persisting Measles Virus in Peripheral Blood Mononuclear Cells From Children With Autism Spectrum Disorder

Yasmin D’Souza, MSca, Eric Fombonne, MDb and Brian J. Ward, MDCMa

a Division of Infectious Diseases, McGill University Health Center, Montreal, Quebec, Canada
b Department of Psychiatry, McGill University, Montreal Children’s Hospital, Montreal, Quebec, Canada

OBJECTIVES. Despite epidemiologic evidence to the contrary, claims of an association between measles-mumps-rubella vaccination and the development of autism have persisted. Such claims are based primarily on the identification of measles virus nucleic acids in tissues and body fluids by polymerase chain reaction. We sought to determine whether measles virus nucleic acids persist in children with autism spectrum disorder compared with control children.

PATIENTS AND METHODS. Peripheral blood mononuclear cells were isolated from 54 children with autism spectrum disorder and 34 developmentally normal children, and up to 4 real-time polymerase chain reaction assays and 2 nested polymerase chain reaction assays were performed. These assays targeted the nucleoprotein, fusion, and hemagglutinin genes of measles virus using previously published primer pairs with detection by SYBR green I. Our own real-time assay targeted the fusion gene using novel primers and an internal fluorescent probe. Positive reactions were evaluated rigorously, and amplicons were sequenced. Finally, anti-measles antibody titers were measured by enzyme immunoassay.

RESULTS. The real-time assays based on previously published primers gave rise to a large number of positive reactions in both autism spectrum disorder and control samples. Almost all of the positive reactions in these assays were eliminated by evaluation of melting curves and amplicon band size. The amplicons for the remaining positive reactions were cloned and sequenced. No sample from either autism spectrum disorder or control groups was found to contain nucleic acids from any measles virus gene. In the nested polymerase chain reaction and in-house assays, none of the samples yielded positive results. Furthermore, there was no difference in anti-measles antibody titers between the autism and control groups.

INTERPRETATION. There is no evidence of measles virus persistence in the peripheral blood mononuclear cells of children with autism spectrum disorder.

Key Words: measles • MMR • autism • autistic spectrum disorder • real-time PCR

Abbreviations: MMR—measles-mumps-rubella • MV—measles virus • ASD—autism spectrum disorder • PCR—polymerase chain reaction • RT—reverse transcription • PBMC—peripheral blood mononuclear cell • IBD—inflammatory bowel disease • HBSS—Hank’s buffered salt solution • GAPDH—glyceraldehyde-3-phosphate dehydrogenase • cDNA—complementary DNA • UNG—uracyl DNA-glycosylase • PBS-T—phosphate-buffered saline-Tween

Accepted Jul 11, 2006.




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