title: 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.
abstract: Specific and sensitive tumor cell detection is becoming increasingly important for diagnosing and staging as well as for the therapeutic management of neuroblastoma patients. We propose a chromogranin A heminested reverse transcription polymerase chain reaction (CgA hn RT-PCR) procedure for the detection of neuroblastoma minimal residual disease in peripheral blood and bone marrow samples. The results were checked in comparison with the presently available procedures (i.e., with the tyrosine hydroxylase nested RT-PCR [TH n RT-PCR] and with the immunocytochemical approach using anti-GD2 antibodies). Controls from healthy patients or from people with unrelated disease (12 samples of bone marrow and 23 samples of peripheral blood) and serial dilution experiments using neuroblastoma cell lines (SKNLP, SKNFI, STA6, STA8) showed CgA hn RT-PCR full specificity and sensitivity ranging from 10(3) to 10(6) (depending on the cell line). The results compared favorably with those obtained using TH n RT-PCR. Preliminary data obtained analyzing bone marrow and peripheral blood specimens from stage IV neuroblastomas showed substantially overlapping results between CgA and TH n RT-PCR procedures. Our data support the potential usefulness of CgA heminested RT-PCR as a specific and sensitive procedure for minimal disease detection in neuroblastoma. A prospective evaluation of this tool in clinical studies might be warranted.
authors.affiliation: Department of Biomedical Sciences and Human Oncology, University of Torino, Italy/ASL10, Pathology Service, Pinerolo, Italy.
authors.forename: Alberto
authors.initials: A
authors.lastname: Pagani
authors.forename: Luigia
authors.initials: L
authors.lastname: Macri
authors.forename: Lawrence B
authors.initials: LB
authors.lastname: Faulkner
authors.forename: Veronica
authors.initials: V
authors.lastname: Tintori
authors.forename: Antonella
authors.initials: A
authors.lastname: Paoli
authors.forename: Alberto
authors.initials: A
authors.lastname: Garaventa
authors.forename: Gianni
authors.initials: G
authors.lastname: Bussolati
mesh_terms: D000293:Adolescent; D000328:Adult; D014408:Biomarkers, Tumor / Q000097:blood / Q000235:genetics; D001853:Bone Marrow / Q000201:enzymology* / Q000473:pathology; D002648:Child; D002675:Child, Preschool; D053379:Chromogranin A; D002864:Chromogranins / Q000097:blood / Q000235:genetics; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D009000:Monocytes / Q000201:enzymology / Q000473:pathology; D017350:N-Acetylgalactosaminyltransferases / Q000097:blood* / Q000276:immunology; D009447:Neuroblastoma / Q000097:blood / Q000201:enzymology* / Q000473:pathology; D012333:RNA, Messenger / Q000032:analysis / Q000097:blood; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D016037:Single-Blind Method; D014407:Tumor Cells, Cultured; D014446:Tyrosine 3-Monooxygenase / Q000097:blood* / Q000235:genetics
chemical_list: D014408:Biomarkers, Tumor; C506150:CHGA protein, human; D053379:Chromogranin A; D002864:Chromogranins; D012333:RNA, Messenger; D014446:Tyrosine 3-Monooxygenase; D017350:N-Acetylgalactosaminyltransferases; C023220:(N-acetylneuraminyl)-galactosylglucosylceramide N-acetylgalactosaminyltransferase