Fill two cell-free DNA (cfDNA) blood collection tubes (STRECK) with 10ml of blood each (20ml total volume; minimum 10ml total with 5ml per tube). Gently invert tubes to mix. Collect two tubes (20 ml total, 10 mL minimum total).
Deliver within 24 hours at room temperature. Do not refrigerate or freeze.
Specimen Preparation
CHOP Staff:STRECK tubes can be obtained from outpatient Buerger and KOP phlebotomy.
External Clients: Please refer to this page to order a Neuroblastoma Liquid Biopsy Kit. Please use tubes by the expiration date printed on the tube.
Kit Contents: 2 STRECK Cell-Free DNA blood collection vacutainers 1 insulated envelope 1 95kPa Bag (i.e. plastic biohazard bag) 1 Aqui-Pak segmented absorbent pouch 1 gel wrap 1 Tray-style transport box 3 Labels: UPS Return, Biohazard, Biological Substance Cat B 2 Forms: Specimen Collection Procedure, DGD Liquid Biopsy Requisition
International Clients: Please refer to this page to order an International Neuroblastoma Liquid Biopsy Kit. Please use tubes by the expiration date printed on the tube.
International Kit Contents: 1 Biotherm 10 Outer Carton (large cardboard outer box) 1 EPS 10 Styrofoam Base and Lid 5 Pharmachill packs (Please store at room temperature. Do NOT refrigerate) 1 Payload carton (small cardboard inner box) 1 Packing Label Sheet (UN3373 label needed from this sheet) 2 STRECK Cell-Free DNA blood collection tubes (BCT) 1 Biohazard bag 3 Forms: Specimen Collection Procedure for International samples, DGD Liquid Biopsy Requisition, and Intelsius Packing Instructions Form
Unacceptable Conditions
The sample is temperature sensitive. Refrigerated or frozen samples will be discarded.
Storage/Transport Temperature
For CHOP Phlebotomy: Samples can be collected throughout the week. Samples collected on weekends or holidays are held in Central Labs and sent to the Genomic Diagnostic Lab the following business day.
For External Clients: Send the sample at room temperature with overnight delivery for receipt Monday through Friday, optimally within 24 hours of collection. A Specimen Collection form is included in the Neuroblastoma kit and can be found here as well. Follow the instructions on the form to collect the sample and ship it back to CHOP.
For International Clients: Send the sample at room temperature for receipt Monday through Friday, optimally within 48-72 hours of collection. A Specimen Collection form is included in the International Neuroblastoma kit and can be found here as well. Follow the instructions on the form to collect the sample and ship it back to CHOP.
If samples need to be held prior to sending to the lab, tubes should be stored vertically.
Volume Required
20ml total (10ml per tube)
Minimum Required
10 ml total
Phlebotomy Draw
After collection, immediately mix by gentle manual inversion 8 to 10 times. Inadequate or delayed mixing may result in incorrect analytical results or poor product performance.
If collecting additional samples from patient during blood draw, Cell-Free DNA blood collection tube should be drawn after EDTA tubes and before any fluoride oxalate (glycolytic inhibitor) tube. If a Cell-Free DNA blood collection tube immediately follows a heparin tube in the draw order, STRECK recommends collecting a nonadditive or EDTA tube as a waste tube prior to collection in the Cell-Free DNA blood collection tube.
Performing Lab
Division of Genomic Diagnostics
Performed
Monday-Friday 9:00am - 4:00pm
Reported
14 days
Utility
Facilitates neuroblastoma disease monitoring, risk stratification, and therapeutic decision-making.
Plasma cfDNA (including ctDNA) is extracted using QIAamp Circulating Nucleic Acid Kit (QIAGEN). The regions of interest are enriched using anchored multiplex PCR method and sequenced using Illumina NovaSeq 6000 system. The sequence data are processed using Archer Analytics and a customized pipeline designed to detect sequence variants in the regions of interest and amplification(s).
This analysis is based on the current understanding of the genomic alterations of neuroblastoma. Clinical implications of some variants may be uncertain or unknown at the time of this report and may change over time. Both somatic and germline variants may be reported and may not be distinguished. If a variant is suspected to be germline, appropriate confirmatory testing should be considered.
The limit of detection for single nucleotide variants and small indels is 0.5% VAF. Manual investigation of certain variants may detect variants between 0.1-0.5% VAF. A negative result does not rule out the presence of an alteration in the patient's plasma that could be due to low ctDNA. The assay may not detect large indels.
The clinical sensitivity of the test will vary due to fluctuations of ctDNA in the plasma through the course of disease and therapy.
This assay is at risk of false negatives when sequence coverage for a ROI is below 800X. In rare situations, some SNVs or indels may not be detectable by this test due to local sequence characteristics or the presence of closely related pseudogenes.
Molecular Testing Notes
Regions of Interest (ROI) (Hg19): ALK, ARID1A, ARID1B, ATRX, BRAF, CDK4, CDKN2A, CDKN2B, CIC, ERBB2, FGFR1 (NM_023110, exons 9-19), HRAS, KRAS, MDM2, MET, MYCN (NM_005378, partial exon 3 for copy number analysis only), NF1, NRAS, PHOX2B, PTPN11, SMARCA4 (NM_003072 exons 17-20, 25 and 26), TERT (NM_198253, promoter region), and TP53. All exons of the genes listed and the flanking intronic sequences are included unless otherwise specified.
CPT Codes
81462
Billing (EAP) Codes
808644
Collection
Collect
Fill two cell-free DNA (cfDNA) blood collection tubes (STRECK) with 10ml of blood each (20ml total volume; minimum 10ml total with 5ml per tube). Gently invert tubes to mix. Collect two tubes (20 ml total, 10 mL minimum total).
Deliver within 24 hours at room temperature. Do not refrigerate or freeze.
Specimen Preparation
CHOP Staff:STRECK tubes can be obtained from outpatient Buerger and KOP phlebotomy.
External Clients: Please refer to this page to order a Neuroblastoma Liquid Biopsy Kit. Please use tubes by the expiration date printed on the tube.
Kit Contents: 2 STRECK Cell-Free DNA blood collection vacutainers 1 insulated envelope 1 95kPa Bag (i.e. plastic biohazard bag) 1 Aqui-Pak segmented absorbent pouch 1 gel wrap 1 Tray-style transport box 3 Labels: UPS Return, Biohazard, Biological Substance Cat B 2 Forms: Specimen Collection Procedure, DGD Liquid Biopsy Requisition
International Clients: Please refer to this page to order an International Neuroblastoma Liquid Biopsy Kit. Please use tubes by the expiration date printed on the tube.
International Kit Contents: 1 Biotherm 10 Outer Carton (large cardboard outer box) 1 EPS 10 Styrofoam Base and Lid 5 Pharmachill packs (Please store at room temperature. Do NOT refrigerate) 1 Payload carton (small cardboard inner box) 1 Packing Label Sheet (UN3373 label needed from this sheet) 2 STRECK Cell-Free DNA blood collection tubes (BCT) 1 Biohazard bag 3 Forms: Specimen Collection Procedure for International samples, DGD Liquid Biopsy Requisition, and Intelsius Packing Instructions Form
Unacceptable Conditions
The sample is temperature sensitive. Refrigerated or frozen samples will be discarded.
Storage/Transport Temperature
For CHOP Phlebotomy: Samples can be collected throughout the week. Samples collected on weekends or holidays are held in Central Labs and sent to the Genomic Diagnostic Lab the following business day.
For External Clients: Send the sample at room temperature with overnight delivery for receipt Monday through Friday, optimally within 24 hours of collection. A Specimen Collection form is included in the Neuroblastoma kit and can be found here as well. Follow the instructions on the form to collect the sample and ship it back to CHOP.
For International Clients: Send the sample at room temperature for receipt Monday through Friday, optimally within 48-72 hours of collection. A Specimen Collection form is included in the International Neuroblastoma kit and can be found here as well. Follow the instructions on the form to collect the sample and ship it back to CHOP.
If samples need to be held prior to sending to the lab, tubes should be stored vertically.
Volume Required
20ml total (10ml per tube)
Minimum Required
10 ml total
Phlebotomy Draw
After collection, immediately mix by gentle manual inversion 8 to 10 times. Inadequate or delayed mixing may result in incorrect analytical results or poor product performance.
If collecting additional samples from patient during blood draw, Cell-Free DNA blood collection tube should be drawn after EDTA tubes and before any fluoride oxalate (glycolytic inhibitor) tube. If a Cell-Free DNA blood collection tube immediately follows a heparin tube in the draw order, STRECK recommends collecting a nonadditive or EDTA tube as a waste tube prior to collection in the Cell-Free DNA blood collection tube.
Ordering
Performing Lab
Division of Genomic Diagnostics
Performed
Monday-Friday 9:00am - 4:00pm
Reported
14 days
Utility
Facilitates neuroblastoma disease monitoring, risk stratification, and therapeutic decision-making.
Plasma cfDNA (including ctDNA) is extracted using QIAamp Circulating Nucleic Acid Kit (QIAGEN). The regions of interest are enriched using anchored multiplex PCR method and sequenced using Illumina NovaSeq 6000 system. The sequence data are processed using Archer Analytics and a customized pipeline designed to detect sequence variants in the regions of interest and amplification(s).
This analysis is based on the current understanding of the genomic alterations of neuroblastoma. Clinical implications of some variants may be uncertain or unknown at the time of this report and may change over time. Both somatic and germline variants may be reported and may not be distinguished. If a variant is suspected to be germline, appropriate confirmatory testing should be considered.
The limit of detection for single nucleotide variants and small indels is 0.5% VAF. Manual investigation of certain variants may detect variants between 0.1-0.5% VAF. A negative result does not rule out the presence of an alteration in the patient's plasma that could be due to low ctDNA. The assay may not detect large indels.
The clinical sensitivity of the test will vary due to fluctuations of ctDNA in the plasma through the course of disease and therapy.
This assay is at risk of false negatives when sequence coverage for a ROI is below 800X. In rare situations, some SNVs or indels may not be detectable by this test due to local sequence characteristics or the presence of closely related pseudogenes.
Molecular Testing Notes
Regions of Interest (ROI) (Hg19): ALK, ARID1A, ARID1B, ATRX, BRAF, CDK4, CDKN2A, CDKN2B, CIC, ERBB2, FGFR1 (NM_023110, exons 9-19), HRAS, KRAS, MDM2, MET, MYCN (NM_005378, partial exon 3 for copy number analysis only), NF1, NRAS, PHOX2B, PTPN11, SMARCA4 (NM_003072 exons 17-20, 25 and 26), TERT (NM_198253, promoter region), and TP53. All exons of the genes listed and the flanking intronic sequences are included unless otherwise specified.
