No evidence of persisting measles virus in peripheral blood mononuclear cells from children with autism spectrum disorder

Yasmin D'Souza, Eric Fombonne, Brian J. Ward

Research output: Contribution to journalArticle

49 Citations (Scopus)

Abstract

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, antimeasles 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 inhouse 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.

Original languageEnglish (US)
Pages (from-to)1664-1675
Number of pages12
JournalPediatrics
Volume118
Issue number4
DOIs
StatePublished - Oct 2006
Externally publishedYes

Fingerprint

Measles virus
Blood Cells
Nucleic Acids
Gene Fusion
Measles
Autistic Disorder
Polymerase Chain Reaction
Mumps
Control Groups
Nucleoproteins
Rubella
Hemagglutinins
Body Fluids
Immunoenzyme Techniques
Fluorescent Dyes
Freezing
Real-Time Polymerase Chain Reaction
Anti-Idiotypic Antibodies
Vaccination
Autism Spectrum Disorder

Keywords

  • Autism
  • Autistic spectrum disorder
  • Measles
  • MMR
  • Real-time PCR

ASJC Scopus subject areas

  • Pediatrics, Perinatology, and Child Health

Cite this

No evidence of persisting measles virus in peripheral blood mononuclear cells from children with autism spectrum disorder. / D'Souza, Yasmin; Fombonne, Eric; Ward, Brian J.

In: Pediatrics, Vol. 118, No. 4, 10.2006, p. 1664-1675.

Research output: Contribution to journalArticle

@article{bb88c7f31c584494a4d3fdca15d5e0e2,
title = "No evidence of persisting measles virus in peripheral blood mononuclear cells from children with autism spectrum disorder",
abstract = "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, antimeasles 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 inhouse 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.",
keywords = "Autism, Autistic spectrum disorder, Measles, MMR, Real-time PCR",
author = "Yasmin D'Souza and Eric Fombonne and Ward, {Brian J.}",
year = "2006",
month = "10",
doi = "10.1542/peds.2006-1262",
language = "English (US)",
volume = "118",
pages = "1664--1675",
journal = "Pediatrics",
issn = "0031-4005",
publisher = "American Academy of Pediatrics",
number = "4",

}

TY - JOUR

T1 - No evidence of persisting measles virus in peripheral blood mononuclear cells from children with autism spectrum disorder

AU - D'Souza, Yasmin

AU - Fombonne, Eric

AU - Ward, Brian J.

PY - 2006/10

Y1 - 2006/10

N2 - 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, antimeasles 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 inhouse 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.

AB - 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, antimeasles 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 inhouse 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.

KW - Autism

KW - Autistic spectrum disorder

KW - Measles

KW - MMR

KW - Real-time PCR

UR - http://www.scopus.com/inward/record.url?scp=33750130715&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33750130715&partnerID=8YFLogxK

U2 - 10.1542/peds.2006-1262

DO - 10.1542/peds.2006-1262

M3 - Article

VL - 118

SP - 1664

EP - 1675

JO - Pediatrics

JF - Pediatrics

SN - 0031-4005

IS - 4

ER -