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1 Division of Clinical Chemistry, Department of Biomedicine and Surgery, Linköping University, S-581 85 Linköping, Sweden.
2 Childhood Cancer Research Unit, Department of Woman and Child Health, Karolinska Institutet, Karolinska Hospital, S-171 76 Stockholm, Sweden.
3 Department of Clinical Chemistry, University Hospital, S-581 85 Linköping, Sweden.
aAuthor for correspondence. Fax 46-13-22-32-40; e-mail bertil.kagedal{at}lio.se.
Background: Sensitive monitoring of minimal residual disease may improve the treatment of neuroblastoma in children. To detect and monitor neuroblastoma cells in blood and bone marrow, we developed a quantitative method for the analysis of tyrosine hydroxylase mRNA.
Methods: We used real-time reverse transcription-PCR. The calibrator was constructed from a segment of tyrosine hydroxylase mRNA that included the target. Blood and bone marrow samples from 24 children with neuroblastoma and 1 child with ganglioneuroma were analyzed. Controls were blood samples from the cords of 40 babies, from 58 children 6 months to 15 years of age, and from 34 healthy adults, as well as from 12 children with other diseases.
Results: The detection limit was 
70 transcripts/mL. All 144 blood controls were below this limit. At diagnosis, blood tyrosine hydroxylase mRNA was higher in children with widespread disease (stage 4/4S; n = 6; range, 203–46 000 transcripts/mL) than in patients with localized disease (stages 1–3; n = 6; 
83 transcripts/mL; P = 0.002). Bone marrow from all five children with localized disease had concentrations <72 transcripts/mL, whereas five of six stage 4 patients had increased concentrations (6000–8 000 000 transcripts/mL; P <0.05). In nine children in whom tyrosine hydroxylase mRNA was measured repeatedly, the results corresponded to the clinical course.
Conclusion: Quantitative analysis of tyrosine hydroxylase mRNA in blood and bone marrow is reliable and easy to perform and may be used for upfront staging, prognostic assessment, and treatment monitoring of neuroblastoma.
The following articles in journals at HighWire Press have cited this article:
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  J. Stutterheim, A. Gerritsen, L. Zappeij-Kannegieter, B. Yalcin, R. Dee, M. M. van Noesel, F. Berthold, R. Versteeg, H. N. Caron, C. E. van der Schoot, et al. Detecting Minimal Residual Disease in Neuroblastoma: The Superiority of a Panel of Real-Time Quantitative PCR Markers Clin. Chem., July 1, 2009; 55(7): 1316 - 1326. [Abstract] [Full Text] [PDF]
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 J. Stutterheim, A. Gerritsen, L. Zappeij-Kannegieter, I. Kleijn, R. Dee, L. Hooft, M. M. van Noesel, M. Bierings, F. Berthold, R. Versteeg, et al. PHOX2B Is a Novel and Specific Marker for Minimal Residual Disease Testing in Neuroblastoma J. Clin. Oncol., November 20, 2008; 26(33): 5443 - 5449. [Abstract] [Full Text] [PDF]
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 I. Y. Cheung, Y. Feng, A. Vickers, W. Gerald, and N.-K. V. Cheung Cyclin D1, a Novel Molecular Marker of Minimal Residual Disease, in Metastatic Neuroblastoma J. Mol. Diagn., April 1, 2007; 9(2): 237 - 241. [Abstract] [Full Text] [PDF]
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 M. V. Corrias, R. Haupt, B. Carlini, S. Parodi, L. Rivabella, A. Garaventa, V. Pistoia, and S. Dallorso Peripheral blood stem cell tumor cell contamination and survival of neuroblastoma patients. Clin. Cancer Res., October 1, 2006; 12(19): 5680 - 5685. [Abstract] [Full Text] [PDF]
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N. Berois, E. Blanc, H. Ripoche, X. Mergui, F. Trajtenberg, S. Cantais, M. Barrois, P. Dessen, B. Kagedal, J. Benard, et al. ppGalNAc-T13: A New Molecular Marker of Bone Marrow Involvement in Neuroblastoma Clin. Chem., September 1, 2006; 52(9): 1701 - 1712. [Abstract] [Full Text] [PDF]
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M. Mitsuhashi, S. Tomozawa, K. Endo, and A. Shinagawa Quantification of mRNA in Whole Blood by Assessing Recovery of RNA and Efficiency of cDNA Synthesis Clin. Chem., April 1, 2006; 52(4): 634 - 642. [Abstract] [Full Text] [PDF]
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K. Swerts, B. De Moerloose, C. Dhooge, J. Vandesompele, C. Hoyoux, K. Beiske, Y. Benoit, G. Laureys, and J. Philippe Potential Application of ELAVL4 Real-Time Quantitative Reverse Transcription-PCR for Detection of Disseminated Neuroblastoma Cells Clin. Chem., March 1, 2006; 52(3): 438 - 445. [Abstract] [Full Text] [PDF]
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 S A Burchill Micrometastases in neuroblastoma: are they clinically important? J. Clin. Pathol., January 1, 2004; 57(1): 14 - 20. [Abstract] [Full Text] [PDF]
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