Establishment and study of different real-time polymerase chain reaction assays for the quantification of cells with deletions of chromosome 7.

The Journal of Molecular Diagnostics : JMD
Elia MattarucchiGiovanni Porta

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 neopl...Continue Reading

References

Feb 24, 2001·Journal of Cancer Research and Clinical Oncology·T Burmeister, E Thiel
Feb 16, 2002·Methods : a Companion to Methods in Enzymology·K J Livak, T D Schmittgen
Jan 18, 2003·Leukemia Research·Anna ButturiniRobert Peter Gale
Aug 9, 2003·Nucleic Acids Research·R G Rutledge, C Côté

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