| Post date: 2023/02/7 |
GoCB protocol
The creation of GoCB has been conducted as a research project. As first step, the GoCB protocol and corresponding standard operation procedures (SOP) were developed considering internationally accepted standards and protocols and for different operational aspects of the GoCB, such as the setting up of the GoCB lab, the GoCB field centers, types of samples, sample collection and sample processing methods, sample preservation and this the access policy for samples.
Types of biological samples
The main biological samples collected by the GoCB include blood sample, urine sample, fresh mucosal biopsies by endoscopy, fresh surgically resected tissue samples and formalin fixed paraffin embedded (FFPE) tissue samples. GoCB systematically collects tumor/non-tumor pairs from all cancer patients, if routine diagnostic sample collection is not hampered.
Collection of data
After obtaining informed consent, data collection is performed by using two GoCB-specific data collection forms. The GoCB data collection form 1 (general questionnaire) consists of patients' demographic data, clinical data and tumor characteristics (site of tumor, morphology, behavior and tumor grade); the latter are collected according to the pathology report. The third edition of the International Classification of Diseases for Oncology (ICD-O-3) is used for coding tumor characteristics, including topography, morphology, behavior and grade.
The GoCB data collection form 2 (risk factor questionnaire) covers a broad range of generic risk factors applicable to all cancer types as well as specific ones based on the epidemiological studies on common cancer types in our population, including anthropometric measures, level of education, occupation, smoking behavior, alcohol and opium consumption, diet, oral hygiene habits and medical history, cancer history in family, exposure to X-ray radiation, exposure to pesticides, physical activity and data on reproductive behaviors (only for women).
Collection of biological samples
The GoCB guidelines for the collection of biological samples include detailed information on each type of biological sample collected, including temperature, time and place of preservation of sample in the filed center, condition of transporting the sample to the GoCB lab, as well as name and telephone number of contact persons in the GoCB laboratory. Additional information includes ischemia time (for surgical resected samples), date and time of sample shipment to the GoCB and cool box temperature during sample shipments.
Following the patient’s informed consent, blood samples (9 ml) are collected using Ethylenediamine tetra acetic acid (EDTA) tubes. Blood samples are shipped to the GoCB laboratory within 2 hours after collection using a cool box and ice pack to ensure appropriate temperature condition during shipment. Mid-stream urine samples are collected using specific sterile containers and transported to GoCB laboratory through the same package and transport facility as blood.
Fresh surgical tissue samples (size of 0.5cm*1cm*1cm) and fresh endoscopy tissue samples are collected using containers with RNA-later and stored in room temperature until shipment to the GoCB laboratory. Formalin fixed surgical tissue samples are collected using prefilled containers with 10% buffered formalin and stored in room temperature until shipment to the GoCB laboratory.
Quality of collected samples and data
Upon receiving the biological samples and questionnaires, the quality of data and biological samples are checked by GoCB secretariat staff to ensure adherence to the GoCB guidelines and protocols. An in-house developed quality control checklist allows to check for the completeness and accuracy of data in a consistent manner. If there is any deviation observed, e.g. missing data, errors in filling out the questionnaire, unclear data, inadequate amount of specimen (volume or size) and errors in labeling, the GoCB secretariat staff provide feedback to the field staff and follow up the issue until resolved.
Processing and preservation of biological samples:
GoCB uses high-quality cryotubes (of 0.5, 1.4 and 3 ml) according to the type of biological sample. The tubes are sterile and free from DNase, RNase, DNA, and endotoxins. Each tube has a unique ID, as a laser etched 2-D barcode on the base. A 2-D barcode reader is used to ensure entering the correct sample code into the GoCB software.
Processing and preservation of blood sample: Blood samples are separated and stored in 1 ml aliquots, as different parts including whole blood (red cap), plasma (white cap), Buffy coat (blue cap) and red blood cell (RBC) (gray cap). Urine samples are stored in 1 ml aliquots too. The samples information is registered into the GoCB software through the sample barcode, as well as detailed information on the sample location (freezer, rack, box, place within the box). After registering in the GoCB software, the samples are transferred and stored in the -80°C freezer.
Processing and preservation of fresh tissue samples: After receiving fresh endoscopy and surgical tissue samples in the GoCB laboratory, tissue containers with RNA-later are placed overnight in refrigerator (4°C). On the next day, the tissue samples are removed from the container and RNA-later and transferred into 0.5 ml cryotube (endoscopy sample) or 3 ml cryotube (surgical resected sample) with external thread screw caps. After registering sample information in the GoCB software, the samples are transferred and stored in the -80°C freezer.
Ethical considerations:
Participants must complete an informed consent form before starting sample and data collection. All information about the project including its benefits and risks as well as indications of using the samples, especially possible use of the samples in international research project are explained to the patient. All patients are well informed about their right to refuse or stop his/her participation at any stage of the project. The GoCB secretariat and field staff as well as clinicians are trained to ensure that participation in the GoCB does not interfere with appropriate patient diagnosis or care. The GoCB uses anonymized codes to ensure the protection of individuals' privacy. The GoCB secretariat and staff have limited access to the dataset according to their role and only a representative of the GoCB steering committee is authorized to have full access to the GoCB dataset. Neither physicians nor patients are reimbursed for participation in the GoCB.
The creation of GoCB has been conducted as a research project. As first step, the GoCB protocol and corresponding standard operation procedures (SOP) were developed considering internationally accepted standards and protocols and for different operational aspects of the GoCB, such as the setting up of the GoCB lab, the GoCB field centers, types of samples, sample collection and sample processing methods, sample preservation and this the access policy for samples.
Types of biological samples
The main biological samples collected by the GoCB include blood sample, urine sample, fresh mucosal biopsies by endoscopy, fresh surgically resected tissue samples and formalin fixed paraffin embedded (FFPE) tissue samples. GoCB systematically collects tumor/non-tumor pairs from all cancer patients, if routine diagnostic sample collection is not hampered.
Collection of data
After obtaining informed consent, data collection is performed by using two GoCB-specific data collection forms. The GoCB data collection form 1 (general questionnaire) consists of patients' demographic data, clinical data and tumor characteristics (site of tumor, morphology, behavior and tumor grade); the latter are collected according to the pathology report. The third edition of the International Classification of Diseases for Oncology (ICD-O-3) is used for coding tumor characteristics, including topography, morphology, behavior and grade.
The GoCB data collection form 2 (risk factor questionnaire) covers a broad range of generic risk factors applicable to all cancer types as well as specific ones based on the epidemiological studies on common cancer types in our population, including anthropometric measures, level of education, occupation, smoking behavior, alcohol and opium consumption, diet, oral hygiene habits and medical history, cancer history in family, exposure to X-ray radiation, exposure to pesticides, physical activity and data on reproductive behaviors (only for women).
Collection of biological samples
The GoCB guidelines for the collection of biological samples include detailed information on each type of biological sample collected, including temperature, time and place of preservation of sample in the filed center, condition of transporting the sample to the GoCB lab, as well as name and telephone number of contact persons in the GoCB laboratory. Additional information includes ischemia time (for surgical resected samples), date and time of sample shipment to the GoCB and cool box temperature during sample shipments.
Following the patient’s informed consent, blood samples (9 ml) are collected using Ethylenediamine tetra acetic acid (EDTA) tubes. Blood samples are shipped to the GoCB laboratory within 2 hours after collection using a cool box and ice pack to ensure appropriate temperature condition during shipment. Mid-stream urine samples are collected using specific sterile containers and transported to GoCB laboratory through the same package and transport facility as blood.
Fresh surgical tissue samples (size of 0.5cm*1cm*1cm) and fresh endoscopy tissue samples are collected using containers with RNA-later and stored in room temperature until shipment to the GoCB laboratory. Formalin fixed surgical tissue samples are collected using prefilled containers with 10% buffered formalin and stored in room temperature until shipment to the GoCB laboratory.
Quality of collected samples and data
Upon receiving the biological samples and questionnaires, the quality of data and biological samples are checked by GoCB secretariat staff to ensure adherence to the GoCB guidelines and protocols. An in-house developed quality control checklist allows to check for the completeness and accuracy of data in a consistent manner. If there is any deviation observed, e.g. missing data, errors in filling out the questionnaire, unclear data, inadequate amount of specimen (volume or size) and errors in labeling, the GoCB secretariat staff provide feedback to the field staff and follow up the issue until resolved.
Processing and preservation of biological samples:
GoCB uses high-quality cryotubes (of 0.5, 1.4 and 3 ml) according to the type of biological sample. The tubes are sterile and free from DNase, RNase, DNA, and endotoxins. Each tube has a unique ID, as a laser etched 2-D barcode on the base. A 2-D barcode reader is used to ensure entering the correct sample code into the GoCB software.
Processing and preservation of blood sample: Blood samples are separated and stored in 1 ml aliquots, as different parts including whole blood (red cap), plasma (white cap), Buffy coat (blue cap) and red blood cell (RBC) (gray cap). Urine samples are stored in 1 ml aliquots too. The samples information is registered into the GoCB software through the sample barcode, as well as detailed information on the sample location (freezer, rack, box, place within the box). After registering in the GoCB software, the samples are transferred and stored in the -80°C freezer.
Processing and preservation of fresh tissue samples: After receiving fresh endoscopy and surgical tissue samples in the GoCB laboratory, tissue containers with RNA-later are placed overnight in refrigerator (4°C). On the next day, the tissue samples are removed from the container and RNA-later and transferred into 0.5 ml cryotube (endoscopy sample) or 3 ml cryotube (surgical resected sample) with external thread screw caps. After registering sample information in the GoCB software, the samples are transferred and stored in the -80°C freezer.
Ethical considerations:
Participants must complete an informed consent form before starting sample and data collection. All information about the project including its benefits and risks as well as indications of using the samples, especially possible use of the samples in international research project are explained to the patient. All patients are well informed about their right to refuse or stop his/her participation at any stage of the project. The GoCB secretariat and field staff as well as clinicians are trained to ensure that participation in the GoCB does not interfere with appropriate patient diagnosis or care. The GoCB uses anonymized codes to ensure the protection of individuals' privacy. The GoCB secretariat and staff have limited access to the dataset according to their role and only a representative of the GoCB steering committee is authorized to have full access to the GoCB dataset. Neither physicians nor patients are reimbursed for participation in the GoCB.
