Linked Life Data

17335844

Source:http://linkedlifedata.com/resource/pubmed/id/17335844

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PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
1-2
pubmed:dateCreated
2007-3-26
pubmed:abstractText
There are two commonly employed types of bioassays for the detection of neutralizing antibodies (NAbs) against interferon-beta (IFNbeta): the cytopatic effect assay (CPE), and the MxA (myxovirus resistance protein A) protein assay (MPA). This article describes a bioassay based on the real time PCR measurement of mRNA that results from the induction, in cultured human cells, of the MxA gene by IFNbeta. Serum samples from 104 patients with multiple sclerosis (MS) treated with IFNbeta were tested for NAbs using our real time PCR bioassay. NAbs also were measured in the same specimens by the MPA assay and CPE assay. The calibration range of the real time PCR bioassay is 0.125-30 LU/mL. The range of the intra- and inter-assay variations (coefficients of variation in log(10)) were 4.05% (range 0.88%-7.90%) and 4.42% (range 0.31%-9.15%), respectively. Samples of the three commercial preparations of IFNbeta-1a and -1b were measured showing dose-response curves parallel to that of the NIH reference IFNbeta (mean SD at the midpoint of the dose-response curve=5%). In addition, the assay was robust with respect to number of cells plated (i.e., increasing cell densities from 12x10(3)/well to 384x10(3)/well resulted in 3.03% variability in MxA expression normalized with glyceraldehyde-3 phosphate dehydrogenase). NAbs titers measured were closely comparable to those obtained by the MPA [r(spearman)=0.899; 89% of observed agreements; K=0.779] and the CPE [r(spearman)=0.7899); 86%; K=0.729] assays. Despite the obvious disadvantage of cost, when carried out according to quality assurance guidelines for molecular diagnostics the new MxA gene-expression assay (MGA) has significant advantages over the other methods for testing NAbs: it has excellent reliability and reproducibility, and utilizes equipment and methodologies already accessible in many clinical laboratories.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Apr
pubmed:issn
0022-1759
pubmed:author
pubmed:issnType
Print
pubmed:day
10
pubmed:volume
321
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
19-31
pubmed:dateRevised
2007-11-15
pubmed:meshHeading
pubmed-meshheading:17335844-Antibodies, pubmed-meshheading:17335844-Biological Assay, pubmed-meshheading:17335844-Calibration, pubmed-meshheading:17335844-Cell Line, Tumor, pubmed-meshheading:17335844-Cytopathogenic Effect, Viral, pubmed-meshheading:17335844-Dose-Response Relationship, Drug, pubmed-meshheading:17335844-Dose-Response Relationship, Immunologic, pubmed-meshheading:17335844-Encephalomyocarditis virus, pubmed-meshheading:17335844-GTP-Binding Proteins, pubmed-meshheading:17335844-Humans, pubmed-meshheading:17335844-Immunologic Factors, pubmed-meshheading:17335844-Interferon-beta, pubmed-meshheading:17335844-Multiple Sclerosis, pubmed-meshheading:17335844-Neutralization Tests, pubmed-meshheading:17335844-RNA, Messenger, pubmed-meshheading:17335844-Reference Standards, pubmed-meshheading:17335844-Reproducibility of Results, pubmed-meshheading:17335844-Reverse Transcriptase Polymerase Chain Reaction, pubmed-meshheading:17335844-Sensitivity and Specificity, pubmed-meshheading:17335844-Time Factors, pubmed-meshheading:17335844-Up-Regulation
pubmed:year
2007
pubmed:articleTitle
Development and validation of a real time PCR-based bioassay for quantification of neutralizing antibodies against human interferon-beta.
pubmed:affiliation
Centro Riferimento Regionale Sclerosi Multipla (CRESM) & Neurobiologia Clinica, Ospedale Universitario S. Luigi Gonzaga, Regione Gonzole 10, I-10043, Orbassano, Torino, Italy. sclerosi.multipla@sanluigi.piemonte.it
pubmed:publicationType
Journal Article, Comparative Study, Research Support, Non-U.S. Gov't, Validation Studies