title: Establishment and study of different real-time polymerase chain reaction assays for the quantification of cells with deletions of chromosome 7.
abstract: The evaluation of residual disease, which has prognostic value in the treatment of hematological malignancies, is currently assessed by scoring a limited number of cells by karyotyping and molecular cytogenetics. Real-time polymerase chain reaction (PCR) is an easier and more sensitive technique, enables analysis of a larger number of cells, and decreases sampling error. However, real-time PCR has been applied only to target transcripts of fusion genes. Here, we considered two real-time PCR strategies to quantify a number of cells carrying a partial deletion of chromosome 7 mixed with normal disomic cells. The first strategy was based on the amplification of two sequences, one on chromosome 7 and the other on chromosome 14. In the second strategy residual disease was assessed by the ratio between the two alleles of a bi-allelic marker, mapped on chromosome 7, measured with allele-specific assays. Precision and accuracy of the two approaches were tested by reference samples with nominal values of residual disease ranging from 2 to 95%. As expected the second strategy resulted in more precise and accurate monitoring within the range from 5 to 95%. Furthermore, this method may be applied to assess the number of dysplastic or neoplastic clones carrying any unbalanced chromosome changes.
authors.affiliation: Department of Experimental and Clinical Biomedical Sciences, University of Insubria, Via Dunant 5, 21100 Varese, Italy.
authors.forename: Elia
authors.initials: E
authors.lastname: Mattarucchi
authors.forename: Milena
authors.initials: M
authors.lastname: Marsoni
authors.forename: Alberto
authors.initials: A
authors.lastname: Passi
authors.forename: Francesco
authors.initials: F
authors.lastname: Lo Curto
authors.forename: Francesco
authors.initials: F
authors.lastname: Pasquali
authors.forename: Giovanni
authors.initials: G
authors.lastname: Porta
mesh_terms: D001483:Base Sequence; D002872:Chromosome Deletion; D002897:Chromosomes, Human, Pair 7 / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D013997:Time Factors