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JMD 2005, Vol. 7, No. 1
Copyright © 2005 American Society for Investigative Pathology & Association for Molecular Pathology

Detection of Central Nervous System Leukemia in Children with Acute Lymphoblastic Leukemia by Real-Time Polymerase Chain Reaction

Sharon R. Pine*, Changhong Yin*, Yousif H. Matloub{dagger}, Hatem E. Sabaawy{ddagger}, Claudio Sandoval*, Oya Levendoglu-Tugal*, M. Fevzi Ozkaynak* and Somasundaram Jayabose*

From the Department of Pediatrics Hematology/Oncology, * New York Medical College, Valhalla, New York; the Department of Pediatrics Hematology/Oncology, {dagger} University of Wisconsin, Madison, Wisconsin; and the Experimental Transplantation and Immunology Branch, {ddagger} Center for Cancer Research, National Cancer Institute, Bethesda, Maryland

Accurate detection of central nervous system (CNS) involvement in children with newly diagnosed acute lymphoblastic leukemia (ALL) could have profound prognostic and therapeutic implications. We examined various cerebrospinal fluid (CSF) preservation methods to yield adequate DNA stability for polymerase chain reaction (PCR) analysis and developed a quantitative real-time PCR assay to detect occult CNS leukemia. Sixty CSF specimens were maintained in several storage conditions for varying amounts of time, and we found that preserving CSF in 1:1 serum-free RPMI tissue culture medium offers the best stability of DNA for PCR analysis. Sixty CSF samples (30 at diagnosis and 30 at the end of induction therapy) from 30 children with ALL were tested for CNS leukemic involvement by real-time PCR using patient-specific antigen receptor gene rearrangement primers. Six of thirty patient diagnosis samples were PCR-positive at levels ranging from 0.5 to 66% leukemic blasts in the CSF. Four of these patients had no clinical or cytomorphological evidence of CNS leukemia involvement at that time. All 30 CSF samples drawn at the end of induction therapy were PCR-negative. The data indicate that real-time PCR analysis of CSF is an excellent tool to assess occult CNS leukemia involvement in patients with ALL and can possibly be used to refine CNS status classification.




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[Abstract] [Full Text] [PDF]


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