{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,27]],"date-time":"2025-10-27T10:30:23Z","timestamp":1761561023064},"reference-count":42,"publisher":"Oxford University Press (OUP)","issue":"3","license":[{"start":{"date-parts":[[2006,3,1]],"date-time":"2006-03-01T00:00:00Z","timestamp":1141171200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/journals\/pages\/open_access\/funder_policies\/chorus\/standard_publication_model"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2006,3,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>Background: Reliable detection of neuroblastoma cells in bone marrow (BM) is critical because BM involvement influences staging, risk assessment, and evaluation of therapeutic response in neuroblastoma patients. Standard cytomorphologic examination of BM aspirates is sensitive enough to detect single tumor cells. Consequently, more sensitive and specific detection methods are indispensable.<\/jats:p>\n               <jats:p>Methods: We used real-time quantitative reverse transcription-PCR (QPCR) of the tyrosine hydroxylase (TH), GD2 synthetase (GALGT), and embryonic lethal, abnormal vision, Drosophila-like 4 (ELAVL4) genes to detect disseminated neuroblastoma cells. We assessed assay sensitivity by addition experiments and then analyzed 97 neuroblastic tumor, BM, peripheral blood (PB), or peripheral blood stem cell (PBSC) samples from 30 patients. The QPCR results were compared with those of a standardized immunocytochemical assay.<\/jats:p>\n               <jats:p>Results: The molecular markers were highly expressed in all evaluated tumor samples. In addition, 32%, 11%, and 38% of all BM, PB, and PBSC samples scored positive for TH, GALGT, or ELAVL4, respectively. The TH and ELAVL4 assays could detect 1 neuroblastoma cell in 106 mononuclear cells. By contrast, the GALGT QPCR assay could detect 1 neuroblastoma cell in 104 mononuclear cells. We assessed the potential prognostic value of TH, GALGT, and ELAVL4 QPCR by analyzing subsequent samples from 3 patients with stage 4 disease. Preliminary results indicated that persistence of high ELAVL4 expression has prognostic value.<\/jats:p>\n               <jats:p>Conclusions: ELAVL4 QPCR can be used to detect residual neuroblastoma cells in clinical samples. However, combination of several molecular markers and screening techniques should be considered to ensure reliable detection of rare neuroblastoma cells.<\/jats:p>","DOI":"10.1373\/clinchem.2005.059485","type":"journal-article","created":{"date-parts":[[2005,12,30]],"date-time":"2005-12-30T01:24:05Z","timestamp":1135905845000},"page":"438-445","source":"Crossref","is-referenced-by-count":26,"title":["Potential Application of ELAVL4 Real-Time Quantitative Reverse Transcription-PCR for Detection of Disseminated Neuroblastoma Cells"],"prefix":"10.1093","volume":"52","author":[{"given":"Katrien","family":"Swerts","sequence":"first","affiliation":[{"name":"Department of Pediatric Hematology and Oncology,"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Barbara","family":"De Moerloose","sequence":"first","affiliation":[{"name":"Department of Pediatric Hematology and Oncology,"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Catharina","family":"Dhooge","sequence":"first","affiliation":[{"name":"Department of Pediatric Hematology and Oncology,"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jo","family":"Vandesompele","sequence":"first","affiliation":[{"name":"Center for Medical Genetics, and Department of Clinical Chemistry, Microbiology and Immunology, Ghent University Hospital, Ghent, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Claire","family":"Hoyoux","sequence":"first","affiliation":[{"name":"Department of Pediatric Hematology and Oncology, Ho\u0302pital de la Citadelle, Lie\u0300ge, Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Klaus","family":"Beiske","sequence":"first","affiliation":[{"name":"Department of Pathology, Rikshospitalet, Oslo, Norway"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yves","family":"Benoit","sequence":"first","affiliation":[{"name":"Department of Pediatric Hematology and Oncology,"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Genevie\u0300ve","family":"Laureys","sequence":"first","affiliation":[{"name":"Department of Pediatric Hematology and Oncology,"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jan","family":"Philippe\u0301","sequence":"first","affiliation":[{"name":"Department of Pediatric Hematology and Oncology,"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"286","published-online":{"date-parts":[[2006,3,1]]},"reference":[{"key":"2020022801330128300_R1","unstructured":"Brodeur GM, Castleberry RP. Neuroblastoma. Pizzo PA Poplack DG eds. Principles and practice of pediatric oncology1997:761-797 Lippincott-Raven Philadelphia. ."},{"key":"2020022801330128300_R2","doi-asserted-by":"crossref","unstructured":"Rill DR, Santana VM, Roberts WM, Nilson T, Bowman LC, Krance RA, et al. Direct demonstration that autologous bone-marrow transplantation for solid tumors can return a multiplicity of tumorigenic cells. Blood1994;84:380-383.","DOI":"10.1182\/blood.V84.2.380.bloodjournal842380"},{"key":"2020022801330128300_R3","doi-asserted-by":"crossref","unstructured":"Brenner MK, Rill DR, Moen RC, Krance RA, Mirro J, Anderson WF, et al. Gene-marking to trace origin of relapse after autologous bone marrow transplantation. Lancet1993;341:85-86.","DOI":"10.1016\/0140-6736(93)92560-G"},{"key":"2020022801330128300_R4","doi-asserted-by":"crossref","unstructured":"Deisseroth AB, Zu ZF, Claxton D, Hanania EG, Fu SQ, Ellerson D, et al. Genetic marking shows that Ph+ cells present in autologous transplants of chronic myelogenous leukemia (CML) contribute to relapse after autologous bone marrow in CML. Blood1994;83:3068-3076.","DOI":"10.1182\/blood.V83.10.3068.3068"},{"key":"2020022801330128300_R5","doi-asserted-by":"crossref","unstructured":"Brodeur GM, Seeger RC, Barrett A, Berthold F, Castleberry RP, D\u2019Angio GJ, et al. International criteria for diagnosis, staging, and response to treatment in patients with neuroblastoma. J Clin Oncol1988;6:1874-1881.","DOI":"10.1200\/JCO.1988.6.12.1874"},{"key":"2020022801330128300_R6","doi-asserted-by":"crossref","unstructured":"Mehes G, Luegmayr A, Kornmuller R, Ambros IM, Ladenstein R, Gadner H, et al. Detection of disseminated tumor cells in neuroblastoma\u20143 log improvement in sensitivity by automatic immunofluorescence plus FISH (AIPH) analysis compared with classical bone marrow cytology. Am J Pathol2003;163:393-399.","DOI":"10.1016\/S0002-9440(10)63669-3"},{"key":"2020022801330128300_R7","doi-asserted-by":"crossref","unstructured":"Cheung NKV, Heller G, Kushner BH, Liu CY, Cheung IY. Detection of metastatic neuroblastoma in bone marrow: when is routine marrow histology insensitive?. J Clin Oncol1997;15:2807-2817.","DOI":"10.1200\/JCO.1997.15.8.2807"},{"key":"2020022801330128300_R8","doi-asserted-by":"crossref","unstructured":"Nagai J, Kigasawa H, Tomioka K, Koga N, Nishihira H, Nagao T. Immunocytochemical detection of bone marrow-invasive neuroblastoma cells. Eur J Haematol1994;53:74-77.","DOI":"10.1111\/j.1600-0609.1994.tb01868.x"},{"key":"2020022801330128300_R9","doi-asserted-by":"crossref","unstructured":"Berthold F, Schneider A, Schumacher A, Bosslet K. Detection of minimal disease in bone-marrow of neuroblastoma patients by immunofluorescence. Pediatr Hematol Oncol1989;6:73-83.","DOI":"10.3109\/08880018909034273"},{"key":"2020022801330128300_R10","doi-asserted-by":"crossref","unstructured":"Cheung NKV, Van Hoff DD, Strandjord SE, Coccia PF. Detection of neuroblastoma cells in bone marrow using GD2 specific monoclonal antibodies. J Clin Oncol1986;4:363-369.","DOI":"10.1200\/JCO.1986.4.3.363"},{"key":"2020022801330128300_R11","doi-asserted-by":"crossref","unstructured":"Swerts K, Ambros PF, Brouzes C, Navarro JM, Gross N, Rampling D, et al. Standardization of the immunocytochemical detection of neuroblastoma cells in bone marrow. J Histochem Cytochem2005;53:1433-1440.","DOI":"10.1369\/jhc.5C6661.2005"},{"key":"2020022801330128300_R12","doi-asserted-by":"crossref","unstructured":"Mehes G, Lo\u0308rch T, Ambros PF. Quantitative analysis of disseminated tumor cells in the bone marrow by automated fluorescence image analysis. Cytometry2000;42:357-362.","DOI":"10.1002\/1097-0320(20001215)42:6<357::AID-CYTO1002>3.0.CO;2-6"},{"key":"2020022801330128300_R13","doi-asserted-by":"crossref","unstructured":"Nagai J, Ishida Y, Koga N, Tanaka Y, Ohnuma K, Toyoda Y, et al. A new sensitive and specific combination of CD81\/CD56\/CD45 monoclonal antibodies for detecting circulating neuroblastoma cells in peripheral blood using flow cytometry. J Pediatr Hematol Oncol2000;22:20-26.","DOI":"10.1097\/00043426-200001000-00004"},{"key":"2020022801330128300_R14","doi-asserted-by":"crossref","unstructured":"Komada Y, Zhang XL, Zhou YW, Inaba H, Deguchi T, Azuma E, et al. Flow cytometric analysis of peripheral blood and bone marrow for tumor cells in patients with neuroblastoma. Cancer1998;82:591-599.","DOI":"10.1002\/(SICI)1097-0142(19980201)82:3<591::AID-CNCR23>3.0.CO;2-W"},{"key":"2020022801330128300_R15","doi-asserted-by":"crossref","unstructured":"Warzynski MJ, Graham DM, Axtell RA, Higgins JV, Hammers YA. Flow cytometric immunophenotyping test for staging\/monitoring neuroblastoma patients. Cytometry2002;50:298-304.","DOI":"10.1002\/cyto.10159"},{"key":"2020022801330128300_R16","doi-asserted-by":"crossref","unstructured":"Tsang KS, Li CK, Tsoi WC, Leung Y, Shing MMK, Chik KW, et al. Detection of micrometastasis of neuroblastoma to bone marrow and tumor dissemination to hematopoietic autografts using flow cytometry and reverse transcriptase-polymerase chain reaction. Cancer2003;97:2887-2897.","DOI":"10.1002\/cncr.11389"},{"key":"2020022801330128300_R17","doi-asserted-by":"crossref","unstructured":"Swerts K, De Moerloose B, Dhooge C, Brichard B, Benoit Y, Laureys G, et al. Detection of residual neuroblastoma cells in bone marrow: comparison of flow cytometry with immunocytochemistry. Cytometry B Clin Cytom2004;1:9-19.","DOI":"10.1002\/cyto.b.20019"},{"key":"2020022801330128300_R18","doi-asserted-by":"crossref","unstructured":"Burchill SA, Bradbury FM, Smith BM, Lewis I, Selby P. Neuroblastoma cell detection by reverse transcriptase-polymerase chain reaction (RT-PCR) for tyrosine hydroxylase mRNA. Int J Cancer1994;57:671-675.","DOI":"10.1002\/ijc.2910570510"},{"key":"2020022801330128300_R19","doi-asserted-by":"crossref","unstructured":"Miyajima Y, Kato K, Numata S, Kudo K, Horibe K. Detection of neuroblastoma cells in bone marrow and peripheral blood at diagnosis by the reverse transcriptase-polymerase chain reaction for tyrosine hydroxylase mRNA. Cancer1995;75:2757-2761.","DOI":"10.1002\/1097-0142(19950601)75:11<2757::AID-CNCR2820751120>3.0.CO;2-S"},{"key":"2020022801330128300_R20","doi-asserted-by":"crossref","unstructured":"Tra\u0308ger C, Kogner P, Lindskog M, Ponthan F, Kullman A, Kagedal B. Quantitative analysis of tyrosine hydroxylase mRNA for sensitive detection of neuroblastoma cells in blood and bone marrow. Clin Chem2003;49:104-112.","DOI":"10.1373\/49.1.104"},{"key":"2020022801330128300_R21","unstructured":"Cheung IY, Cheung NKV. Molecular detection of GAGE expression in peripheral blood and bone marrow: utility as a tumor marker for neuroblastoma. Clin Cancer Res1997;3:821-826."},{"key":"2020022801330128300_R22","doi-asserted-by":"crossref","unstructured":"Oltra S, Martinez F, Orellana C, Grau E, Fernandez JM, Canete A, et al. Minimal residual disease in neuroblastoma: to GAGE or not to GAGE. Oncol Res2004;14:291-295.","DOI":"10.3727\/096504003773994824"},{"key":"2020022801330128300_R23","unstructured":"Cheung IY, Barber D, Cheung NKV. Detection of microscopic neuroblastoma in marrow by histology, immunocytology, and reverse transcription-PCR of multiple molecular markers. Clin Cancer Res1998;4:2801-2805."},{"key":"2020022801330128300_R24","doi-asserted-by":"crossref","unstructured":"Gilbert J, Norris MD, Marshall GM, Haber M. Low specificity of PGP9.5 expression for detection of micrometastatic neuroblastoma. Brit J Cancer1997;75:1779-1781.","DOI":"10.1038\/bjc.1997.303"},{"key":"2020022801330128300_R25","unstructured":"Mattano LA, Moss TJ, Emerson SG. Sensitive detection of rare circulating neuroblastoma cells by reverse transcriptase-polymerase chain reaction. Cancer Res1992;52:4701-4705."},{"key":"2020022801330128300_R26","unstructured":"Cheung IY, Cheung NKV. Quantitation of marrow disease in neuroblastoma by real-time reverse transcription-PCR. Clin Cancer Res2001;7:1698-1705."},{"key":"2020022801330128300_R27","doi-asserted-by":"crossref","unstructured":"Hoon DSB, Kuo CT, Wen S, Wang H, Metelista L, Reynolds CP, et al. Ganglioside GM2\/GD2 synthetase mRNA is a marker for detection of infrequent neuroblastoma cells in bone marrow. Am J Pathol2001;159:493-500.","DOI":"10.1016\/S0002-9440(10)61721-X"},{"key":"2020022801330128300_R28","doi-asserted-by":"crossref","unstructured":"Bozzi F, Luksch R, Collini P, Gambirasio F, Barzano\u0300 E, Polastri D, et al. Molecular detection of dopamine decarboxylase expression by means of reverse transcriptase and polymerase chain reaction in bone marrow and peripheral blood: utility as a tumor marker for neuroblastoma. Diagn Mol Pathol2004;13:135-143.","DOI":"10.1097\/01.pdm.0000128699.14504.06"},{"key":"2020022801330128300_R29","doi-asserted-by":"crossref","unstructured":"Pagani A, Macri L, Faulkner LB, Tintori V, Paoli A, Garaventa A, et al. Detection procedures for neuroblastoma cells metastatic to blood and bone marrow: blinded comparison of chromogranin A heminested reverse transcription polymerase chain reaction to tyrosine hydroxylase nested reverse transcription polymerase chain reaction and to anti-GD2 immunocytology. Diagn Mol Pathol2002;11:98-106.","DOI":"10.1097\/00019606-200206000-00006"},{"key":"2020022801330128300_R30","doi-asserted-by":"crossref","unstructured":"Oltra S, Martinez F, Orellana C, Grau E, Fernandez JM, Canete A, et al. The doublecortin gene, a new molecular marker to detect minimal residual disease in neuroblastoma. Diagn Mol Pathol2005;14:53-57.","DOI":"10.1097\/01.pas.0000149876.32376.c0"},{"key":"2020022801330128300_R31","doi-asserted-by":"crossref","unstructured":"Furukawa K, Soejima H, Niikawa N, Shiku H, Furukawa K. Genomic organization and chromosomal assignment of the human \u03b21,4-N-acetylgalactosaminyltransferase gene. J Biol Chem1996;271:20836-20844.","DOI":"10.1074\/jbc.271.34.20836"},{"key":"2020022801330128300_R32","unstructured":"Ruan S, Lloyd KO. Glycosylation pathways in the biosynthesis of gangliosides in melanoma and neuroblastoma cells: relative glycosyltransferase levels determine ganglioside patterns. Cancer Res1992;52:5725-5731."},{"key":"2020022801330128300_R33","unstructured":"Dalmau J, Furneaux HM, Cordon-Cardo C, Posner JB. The expression of the Hu (paraneoplastic encephalomyelitis\/sensory neuronopathy) antigen in human normal and tumor tissues. Am J Pathol1992;141:881-886."},{"key":"2020022801330128300_R34","doi-asserted-by":"crossref","unstructured":"Ambros IM, Benard J, Boavida M, Bown N, Caron H, Combaret V, et al. Quality assessment of genetic markers used for therapy stratification. J Clin Oncol2003;21:2077-2084.","DOI":"10.1200\/JCO.2003.03.025"},{"key":"2020022801330128300_R35","doi-asserted-by":"crossref","unstructured":"Vandesompele J, De Preter K, Pattyn F, Poppe B, Van Roy N, De Paepe A, et al. Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol2002;3:research0034.1-0034.11.","DOI":"10.1186\/gb-2002-3-7-research0034"},{"key":"2020022801330128300_R36","doi-asserted-by":"crossref","unstructured":"Beillard E, Pallisgaard N, van der Velden VHJ, Bi W, Dee R, van der Schoot E, et al. Evaluation of candidate control genes for diagnosis and residual disease detection in leukemic patients using \u2018real-time\u2019 quantitative reverse-transcriptase polymerase chain reaction (RQ-PCR)\u2014a Europe against Cancer program. Leukemia2003;17:2474-2486.","DOI":"10.1038\/sj.leu.2403136"},{"key":"2020022801330128300_R37","doi-asserted-by":"crossref","unstructured":"Hartmann O, Valteau-Couanet D, Vassal G, Lapierre V, Brugie\u0300res L, Delgado R, et al. Prognostic factors in metastatic neuroblastoma in patients over 1 year of age treated with high-dose chemotherapy and stem cell transplantation: a multivariate analysis in 218 patients treated in a single institution. Bone Marrow Transplant1999;23:789-795.","DOI":"10.1038\/sj.bmt.1701737"},{"key":"2020022801330128300_R38","doi-asserted-by":"crossref","unstructured":"Cotterill SJ, Pearson ADJ, Pritchard J, Foot ABM, Roald B, Kohler JA, et al. Clinical prognostic factors in 1277 patients with neuroblastoma: results of The European Neuroblastoma Study Group \u2018Survey\u2019 1982\u20131992. Eur J Cancer2000;36:901-908.","DOI":"10.1016\/S0959-8049(00)00058-7"},{"key":"2020022801330128300_R39","doi-asserted-by":"crossref","unstructured":"Faulkner LB, Garaventa A, Paoli A, Tintori V, Tamburini A, Lacitignola L, et al. In vitro cytoreduction studies and cell sorting-enhanced tumor-cell detection in high-risk neuroblastoma patients: implications for leukapheresis strategies. J Clin Oncol2000;18:3829-3836.","DOI":"10.1200\/JCO.2000.18.22.3829"},{"key":"2020022801330128300_R40","unstructured":"Lambooy LHJ, Gidding CEM, van den Heuvel LP, Hulsbergen-van de Kaa CA, Ligtenberg M, Bo\u0308kkerink JPM, et al. Real-time analysis of tyrosine hydroxylase gene expression: a sensitive and semiquantitative marker for minimal residual disease detection in neuroblastoma. Clin Cancer Res2003;9:812-819."},{"key":"2020022801330128300_R41","doi-asserted-by":"crossref","unstructured":"Cheung IY, Lo Piccolo MS, Kushner BH, Kramer K, Cheung NKV. Quantitation of GD2 synthase mRNA by real-time reverse transcriptase polymerase chain reaction: clinical utility in evaluating adjuvant therapy in neuroblastoma. J Clin Oncol2003;21:1087-1093.","DOI":"10.1200\/JCO.2003.02.055"},{"key":"2020022801330128300_R42","doi-asserted-by":"crossref","unstructured":"Cheung IY, Lo Piccolo MS, Kushner BH, Cheung NKV. Early molecular response of marrow disease to biologic therapy is highly prognostic in neuroblastoma. J Clin Oncol2003;21:3853-3858.","DOI":"10.1200\/JCO.2003.11.077"}],"container-title":["Clinical Chemistry"],"original-title":[],"language":"en","link":[{"URL":"http:\/\/academic.oup.com\/clinchem\/article-pdf\/52\/3\/438\/32686675\/clinchem0438.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"http:\/\/academic.oup.com\/clinchem\/article-pdf\/52\/3\/438\/32686675\/clinchem0438.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2020,2,28]],"date-time":"2020-02-28T06:36:57Z","timestamp":1582871817000},"score":1,"resource":{"primary":{"URL":"https:\/\/academic.oup.com\/clinchem\/article\/52\/3\/438\/5626699"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2006,3,1]]},"references-count":42,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2006,3,1]]},"published-print":{"date-parts":[[2006,3,1]]}},"URL":"https:\/\/doi.org\/10.1373\/clinchem.2005.059485","relation":{},"ISSN":["0009-9147","1530-8561"],"issn-type":[{"value":"0009-9147","type":"print"},{"value":"1530-8561","type":"electronic"}],"subject":[],"published-other":{"date-parts":[[2006,3,1]]},"published":{"date-parts":[[2006,3,1]]}}}