Fast molecular diagnostics of canine T-cell lymphoma by PCR and capillary gel electrophoresis with laser-induced fluorescence detector

J Chromatogr B Analyt Technol Biomed Life Sci. 2007 Jul 1;854(1-2):268-72. doi: 10.1016/j.jchromb.2007.04.050. Epub 2007 May 17.

Abstract

Lymphoma is the most common hematopoietic tumor in dogs and manifests as a proliferation of malignant lymphoid cells primarily affecting the lymph nodes or solid visceral organs. We describe the use of capillary gel electrophoresis (CGE) with a laser-induced fluorescence (LIF) detector based on polymerase chain reaction (PCR) to rapidly detect a disorder of the canine T-cell receptor gamma (TCRgamma) gene. After the PCR amplification of the specific TCR( gene in dogs, the 90-bp DNA fragment amplified was separated in a fused-silica capillary by CGE-LIF. Under an electric field of 375 V/cm and with a sieving matrix of 1.5% poly (ethyleneoxide) (M(r) 600,000), the amplified PCR products were analyzed within 4 min by CGE separation. When the CGE-LIF method was applied to real clinical samples of the specific DNA fragment of the TCR( gene, the migration time and the corrected peak area showed relative standard deviations (n=5) of 0.29% and 0.58%, respectively. Both methods of CGE-LIF and slab gel electrophoresis showed same results for nine clinical samples. This PCR/CGE-LIF technique may prove to be a new fast and simple tool for the rapid diagnosis of the PCR-amplified DNA of canine T-cell lymphoma.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • DNA Primers
  • Dogs
  • Electrophoresis, Capillary / methods*
  • Lasers
  • Male
  • Polymerase Chain Reaction / methods*

Substances

  • DNA Primers