Detection Evaluation of a Novel Blood-based DNA Methylation Assay in Early-stage Hepatocellular Carcinoma Patients
Conditions: Liver Cancer
Sponsor: Genetron Health

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Detection Evaluation of a Novel Blood-based DNA Methylation 
Assay in Early-stage Hepatocellular Carcinoma Patients 
 
 
 
 
 
 
Study Document 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
NTC number: NCT05343832 
Date: 05/05/2022 
 

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Content 
 
1. 
BACKGROUND ...................................................................................................................................... 4 
2. 
OBJECTIVE OF CLINICAL TRIAL ...................................................................................................... 6 
3. 
OVERALL DESIGN OF CLINICAL TRIAL .......................................................................................... 6 
3.1 SELECTION OF CLINICAL TRIAL CENTERS ................................................................................................. 6 
3.2 TEST PERFORMANCE EVALUATION ........................................................................................................... 7 
3.2.1 Test performance evaluation method ............................................................................................... 7 
3.2.2 Determination of the sample size for test performance evaluation .................................................. 7 
3.2.3 Inclusion Criteria ............................................................................................................................. 7 
3.2.4 Exclusion Criteria ............................................................................................................................ 8 
3.3 DIAGNOSTIC PERFORMANCE EVALUATION ............................................................................................... 8 
3.3.1 
Diagnostic performance evaluation method................................................................................. 8 
3.3.2 
Sample size for diagnostic performance evaluation ..................................................................... 9 
3.3.3 
Inclusion Criteria ......................................................................................................................... 9 
3.3.4 
Exclusion Criteria ........................................................................................................................ 9 
3.3.5 
Judgment criteria for negative and positive result of test reagent ................................................ 9 
3.4 
SAMPLE REQUIREMENTS ................................................................................................................. 10 
3.4.1 
Sample collection ....................................................................................................................... 10 
3.4.2 
DNA extraction and sample testing ........................................................................................... 10 
3.4.3 
Storage and transportation ......................................................................................................... 10 
3.5 
TRIAL IMPLEMENTATION PROCESS .................................................................................................. 11 
3.5.1 
Number of included samples ..................................................................................................... 11 
3.5.2 
Blinding ..................................................................................................................................... 11 
3.5.3 
Unblinding ................................................................................................................................. 12 
3.6 
DURATION OF CLINICAL TRIAL ....................................................................................................... 12 
3.7 
STATISTICAL METHOD OF CLINICAL TRIAL ..................................................................................... 12 
3.7.1 
Principles of statistical analysis ................................................................................................. 12 
3.7.2 
Analysis of general data ............................................................................................................. 12 
3.7.3 
Statistical evaluation of test performance .................................................................................. 13 
3.7.4 
Statistical evaluation of diagnostic performance ....................................................................... 13 
3.7.5 
Comparison of sensitivity across different test methods for screening liver cancer .................. 14 
3.8 
QUALITY CONTROL OF CLINICAL TRIALS ......................................................................................... 14 
3.9 
CONTROLS FOR BIAS ....................................................................................................................... 15 
3.10 
DATA PROCESSING AND RECORD KEEPING ....................................................................................... 15 
3.11 
MANAGEMENT OF SAMPLES AND INSTRUMENTS ............................................................................. 16 
3.12 
AUDITING ....................................................................................................................................... 16 
4. 
ETHICAL ISSUES AND INFORMED CONSENT .............................................................................. 16 
4.1 
ETHICAL CONSIDERATIONS ............................................................................................................. 16 
4.2 
INFORMED CONSENT PROCESS AND INFORMED CONSENT TEXT ....................................................... 17 

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5. 
ANALYSIS OF THE POSSIBILITY OF ADVERSE EVENTS, AND PROVISIONS FOR 
REPORTING ADVERSE EVENTS AND PRODUCT DEFECTS ................................................................ 18 
5.1 
ADVERSE EVENT ............................................................................................................................. 18 
5.2 
SERIOUS ADVERSE EVENT ............................................................................................................... 18 
5.3 
DEVICE DEFECT .............................................................................................................................. 19 
6. 
PROVISIONS ON THE REVISION OF CLINICAL TRIAL PROTOCOLS ....................................... 20 
7. 
WHAT SHOULD BE COVERED IN A CLINICAL TRIAL REPORT ................................................. 20 
8. 
CONFIDENTIALITY ............................................................................................................................ 20 
9. 
RESPONSIBILITIES OF EACH PARTY .............................................................................................. 21 
9.1 
RESPONSIBILITIES OF THE SPONSOR ................................................................................................ 21 
9.2 
RESPONSIBILITIES OF CLINICAL TRIAL INSTITUTIONS ...................................................................... 21 
 
 
 
 

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1. Background 
Primary liver malignancy (primary liver cancer) occurs at the epithelial or 
mesenchymal tissue of the liver, mainly including hepatocellular carcinoma, intrahepatic 
cholangiocarcinoma and combined hepatocellular cholangiocarcinoma, in which 
hepatocellular carcinoma accounts for more than 85%~90%, being the most common 
malignant tumor in China. 
Currently, the incidence of liver cancer is steadily increasing at a rate of 3% per year 
around the world. The Asia and China have the highest burden of liver cancer, 70% of 
patients with newly-developed liver cancer is from Asia and 50% from China. 
Correspondingly, the number of patients died from liver cancer in China also accounts for 
half of that in the world. Chinese Cancer Center published the latest data about the cancer 
on Chinese Journal of Oncology in January 2018: In total number of patients with 
newly-developed malignant tumors in China, the frequency of liver cancer ranks fourth 
(365,000 cases/year); In total number of patients died from malignant tumors, the 
frequency of liver cancer ranks second (319,000 cases/year). The risk of liver cancer is 
especially higher in men older than 40 years old. 
The pathogenesis and exact molecular mechanism of primary liver cancer are not yet 
fully understood. At present, the pathogenesis of primary liver cancer is believed to be a 
complex process with multiple factors and steps, which is affected by both environment 
and diet. The data of epidemiology and relevant studies show that infections of hepatitis B 
virus (HBV) and hepatitis C virus (HCV), aflatoxin, water pollution, alcohol, liver 
cirrhosis, sex hormones, nitrosamines, trace elements are associated with liver cancer. 
Secondary liver cancer (metastatic liver cancer) can induce the disease through different 
pathways, such as blood, lymphatic metastasis, or direct invasion of the liver. 
At present, the routine surveillance and screening indicators for the early diagnosis of 
liver cancer mainly include alpha-fetoprotein (AFP) and liver ultrasound examination 
(US). Based on domestic and foreign reports and clinical verification, the sensitivity of 

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AFP in diagnosing liver cancer is about 60%, the false negative rate is 40%, the specificity 
is about 80%, and the false positive rate is 20%. For these reasons, AFP is no longer a 
screening indicator in AASLD guideline. For nodules larger than 2 cm, the sensitivity of 
US is still 84%; but for nodules of 0.5~2 cm in the liver, the sensitivity is only 33%. 
Therefore, there is an urgent need to find out a new biomarker for auxiliary diagnosis of 
liver cancer with higher sensitivity and specificity compared with current screening 
methods. 
ctDNA, also known as Cell-free tumor DNA or circulating tumor DNA, circulates 
freely in the blood of patients with cancer. The analysis and comparison between ctDNA 
mutation and the site of normal genome by means of invasive approach can provide the 
useful information for tumor diagnosis, prognosis, and monitoring. 
ctDNA generates from necrotic or apoptotic tumor cells, circulating tumor cells, and 
exosomes secreted by tumor cells. In the body of cancer patients, the number of ctDNA is 
small, the half life is short, and the size of ctDNA is about 160~180 bp. These fragments 
can reflect the number of tumor cells and its change. ctDNA is the "identity fingerprint" 
released by tumor cells into the blood, which is a tumor marker with high sensitivity and 
specificity . ctDNA is a non-invasive alternative for patients who are not suitable for 
tumor biopsy, which not only relieves the suffering, but also serves as an alternative to 
repeated invasive biopsy. 
DNA methylation refers to the covalent bonding of a methyl group at the 5’ carbon 
position of cytosine of genomic CpG dinucleotides under the action of DNA 
methyltransferases. DNA methylation is an important epigenetic mark information and 
one of the earliest discovered DNA modification pathways. It plays an important role in 
maintaining normal cell function, genetic imprinting, embryonic development and human 
tumorigenesis, and is also one of the popular topics of study. Alterations in DNA 
methylation level and pattern are an important factor in tumorigenesis. In normal cells, the 
CpG island located in the promoter region of tumor suppressor genes is in a low level or 
unmethylated state. At this time, tumor suppressor genes are in a normal open state, and 
the expression of tumor suppressor genes inhibits tumorigenesis. In tumor cells, the CpG 

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island in this region is highly methylated, the chromatin conformation is altered, and 
tumor suppressor gene expression is turned off, resulting in cell entry into the cell cycle, 
loss of apoptosis, defective DNA repair, dysfunction of vessel growth and cellular 
adhesion, eventually leading to tumorigenesis. DNA methylation is an ideal biomarker and 
evaluation indicator for prognosis and early diagnosis of tumors, and it is of great 
significance for tumor screening, risk assessment, early diagnosis, staging, prognosis 
judgment, treatment and monitoring. 
DNA methylation changes are an early event in the tumorigenesis of hepatic cancer, 
including increased methylation of tumor suppressor genes or decreased methylation of 
proto-oncogenes, and this change precedes mutation of driver genes. Therefore, changes in 
methylation patterns are considered to be the first detectable indicator closely related to 
tumorigenesis, providing a screening window for early diagnosis of liver cancer. By 
analyzing the ctDNA methylation model in a large number of patients with liver cancer 
and normal individuals, we finally screened out 6 liver cancer ctDNA-specific gene 
methylation sites, and established a comprehensive methylation diagnostic model with 
high-sensitivity and specificity. On this basis, the human 6 gene methylation test kit 
(PCR-fluorescent probe method), which is a novel blood-based DNA methylation assay, 
was developed. 
2. Objective of Clinical Trial 
1) Test performance evaluation 
2) Diagnostic performance evaluation 
3. Overall Design of Clinical Trial 
3.1 Selection of clinical trial centers 
The representative centers are selected to conduct this clinical trial, based on the 
consideration of the characteristics of the product, the intended use, the demographics, the 
epidemiologic background, the features of pathogenic microorganism, the study 
population, the clinical conditions (expected use environment and users). 

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3.2 Test performance evaluation 
3.2.1 Test performance evaluation method 
Following the principle of concurrent blinding, the blood samples are collected from 
the non-HCC patients and patients with liver cancer who had been diagnosed according to 
the clinical diagnostic criteria for primary liver cancer. The clinical diagnosis of primary 
liver cancer is determined in accordance with the clinical diagnostic criteria in Guidelines 
for diagnosis and treatment of primary liver cancer (2019 Edition) set by the National 
Health Commission. The enrolled subjected are coded. The selected samples are tested 
with test reagent and the control method high-throughput human methylation sequencing 
(NGS sequencing method), the test results are compared. 
3.2.2 Determination of the sample size for test performance evaluation 
According to the pre-clinical independent cohort verification results of 6 liver 
cancer-specific methylation sites that were screened out, the number of positive samples 
should be not less than 78; the number of negative samples should be not less than 238; 
the samples available in this study should be not less than 316 cases. 
Considering that the mutation rate of some genes is extremely low (AK055957, 
DAB2IP, TBX15, GRASP, PPFIA1, PSD4), the subjects should be enrolled sequentially, 
and at least one positive case should be screened out in each type of gene mutation. If the 
requirements are not met, more subjects should be enrolled for testing. It should be 
ensured that all samples are traceable. 
3.2.3 Inclusion Criteria 
1) Confirmed primary hepatocellular carcinoma or confirmed non-HCC; 
2) I or my legal representative can read, understand and sign the informed consent; 
3) Agree to provide blood samples and have good clinical compliance; 
4) The basic clinical information is complete, including: the patient's unique 
traceability number (ID number/outpatient clinic number/medical insurance card number), 
age, gender, imaging and/or pathological diagnosis results (for patients with primary liver 

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cancer), imaging examination confirmed non-identical Liver cancer (non-HCC patients). 
3.2.4 Exclusion Criteria 
1) pregnant women; 
2) Have received an organ transplant; 
3) Non-HCC patients diagnosed with other tumors; 
4) Patients with primary hepatocellular carcinoma combined with other tumors; 
5) The investigator judges that they are not eligible for inclusion. 
3.3 Diagnostic performance evaluation 
3.3.1 Diagnostic performance evaluation method 
According to the clinical diagnosis results of typical primary liver cancer imaging, 
the enrolled subjects at a high risk of primary liver cancer should undergo 
alpha-fetoprotein (AFP) test, liver ultrasound examination (US), test with test reagent and 
dynamic contrast-enhanced MRI examination, and the sensitivity and specificity of the 
test reagent should be calculated. AFP will be tested by the conventional method of each 
center and the method specified by the sponsor. 
According to the clinical diagnostic criteria in Guidelines for diagnosis and treatment 
of primary liver cancer (2019 Edition) set by the National Health Commission: The 
clinical diagnosis of liver cancer can be made, when a nodule with a diameter ≤ 2 cm is 
found, and minimum 2 examination results in dynamic contrast-enhanced MRI, dynamic 
contrast-enhanced CT, ultrasound examination and hepatocyte-specific contrast agent 
Gd-EOB-DTPA contrast MRI show significant brightness during arterial phase, lower 
brightness of lesion than liver parenchyma during portal venous phase and equilibrium 
phase (The concentration of the contrast is significantly higher than that of the normal 
hepatic tumor tissue during arterial phase in a very short period, then decreases during 
portal venous phase. It’s a typical characteristic of liver cancer); The clinical diagnosis of 
liver cancer can be made, when a nodule with a diameter > 2 cm is found. Therefore, for 
subjects with typical imaging manifestations of liver cancer but the diameter of 

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intrahepatic nodule ≤ 2 cm, an additional imaging examination will be performed to 
confirm the diagnosis. 
3.3.2 Sample size for diagnostic performance evaluation 
In this study, the useful samples are planned to be no less than 4500. And each 
clinical trial center should cherry-pick the subjects and avoid serious bias. If the relevant 
regulations and policies change during the clinical trial, the trial protocol should be 
reasonably adjusted to meet the requirements of relevant national regulations and policies, 
if necessary. 
3.3.3 Inclusion Criteria 
 1) Have high risk factors for liver cancer such as HBV and/or HCV infection, 
alcoholic liver disease, non-alcoholic steatohepatitis, long-term consumption of food 
contaminated with aflatoxin, liver cirrhosis caused by various other reasons, and family 
history of liver cancer Wait; 
2) I or my legal representative can read, understand and sign the informed consent; 
3) Agree to provide blood samples, be able to receive imaging examinations and have 
good clinical compliance; 
4) The complete clinical basic information includes: the patient's unique traceability 
number (ID number/outpatient clinic number/medical insurance card number), age, gender, 
etc. 
3.3.4 Exclusion Criteria 
1) pregnant women; 
2) Have received an organ transplant; 
3) Diagnosed with other tumors; 
4) The investigators judged that they were not eligible for inclusion. 
3.3.5 Judgment criteria for negative and positive result of test reagent 
The test results of the test reagent are judged according to the positive value in the 
instructions for use: 

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When the HCCscan value is greater than 58, the sample is judged to be positive; 
when the HCCscan value is less than or equal to 58, the sample is judged to be negative. 
For the examinations of "US+AFP", the negative result and positive result are 
determined according to the following criteria: 
1） The results of "US+AFP" is positive if one of the following situations is met: 
(1) Serum AFP > 400 ng/mL, regardless of whether any nodule is detected by 
ultrasound examination; 
(2) A nodule ≥ 2 cm is detected by ultrasound examination, regardless of AFP 
serum concentration; 
(3) A nodule ≥ 1 cm but < 2 cm is detected by ultrasound examination, and serum 
AFP ≥ 200 ng/mL 
2） Results that do not meet the above criteria are recorded as "US+AFP" negative. 
3.4 Sample requirements 
3.4.1 Sample collection 
1） Test performance evaluation: 20 mL of peripheral venous blood is needed for the 
testing with test reagent and control method (note: 10 mL/tube, in total of two 
tubes). Transfer the blood into a disposable vacuum blood collection tube, mix 
the blood and anticoagulant by gently inverting the tube 5~6 times. 
2） Diagnostic performance evaluation: 10 mL of peripheral venous blood is needed 
(note: 10 mL/tube). Transfer the blood into a disposable vacuum blood collection 
tube, mix the blood and anticoagulant by gently inverting the tube 5~6 times. 
3.4.2 DNA extraction and sample testing 
Samples are processed in strict accordance with standard operating procedures. After 
nucleic acid extraction, samples should be tested with the test reagent and control method 
according to the instructions for use, and the raw data should be kept at the same time. 
3.4.3 Storage and transportation 
Blood samples in disposable blood collection tubes should be stored at 6~37°C for no 
more than 7 days. 

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The separated plasma samples should be subject for extraction immediately. If the 
samples cannot be processed immediately, they should be stored in a -20±5℃ refrigerator 
for no more than 6 months. 
The extracted cfDNA should be stored at -20±5℃ for no more than 6 months. The 
the extracted cfDNA should be transported upon dry ice and the transportation period 
should not be more than 110 hours. 
3.5 Trial Implementation Process 
3.5.1 Number of included samples 
The included samples are numbered before testing, and the numbers are recorded in 
Screening Inclusion Form. Each clinical trial center will appoint a designated person 
responsible for numbering the sample that meet the inclusion criteria. 
Principle of sample numbering: 
Each sample has a 1 letter (A/B, group number) plus a unique 5-digit number. 
A represents the group for performance evaluation of test reagent, and B represents 
the group for percent agreement evaluation of test reagent. 
The 1st digit of the number represents the code of the clinical trial center, and the 2nd 
to 5th digits represent the sequence of inclusion: 
For example: the first included sample from the Third Affiliated Hospital of Sun 
Yat-sen University in the group for performance evaluation: A10001. 
A  1 0001 
Inclusion sequence number 
             The code of the clinical trial center 
             Test group number 
3.5.2 Blinding 
In principle, one subject only provides one sample, and each subject should have a 
unique clinical trial number (secondary code). After the samples are included and before 
the test implementation, the blinding personnel should stick the secondary number on the 
tube, and save the mapping table of the secondary number and primary number (the 

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unique traceable number of the subject). Before sample testing, the Blindness Record 
should be developed by an investigator and reviewed by another investigator for both of 
them who do not participate in sample testing, then retained by the blinding personnel 
independently. During the test, only the secondary code of the sample is knowable. 
3.5.3 Unblinding 
After the samples are tested with the test reagent and control method, the results are 
unblinded. 
3.6 Duration of Clinical Trial 
This clinical trial went through protocol drafting, project establishment, ethics review, 
agreement signing, human genetic resources approval, registration and back-up, clinical 
initiation and pre-test, enrollment, completion of enrollment, statistical analysis, 
completion of report, and seal, the duration is about 18 ~ 36 months. If the sample 
collection is smooth and the number of samples meets the test requirements, the duration 
clinical trial can be shortened; if it is not smooth, the duration can be appropriately 
extended. 
3.7 Statistical Method of Clinical Trial 
3.7.1 Principles of statistical analysis 
SAS9.4 software (or higher version) is used for statistical analysis, and PASS16 
software (or higher version) is used for sample size calculation. All statistical tests are 
two-sided (unless otherwise stated). 
Basic principles of statistical description: The quantitative data (such as age, gender, 
etc.) are expressed in forms of mean, standard deviation, median, minimum value, 
maximum value, lower quartile (Q1), and upper quartile (Q3) according to the distribution 
of the data; the qualitative data are expressed in forms of the number of cases and the 
corresponding proportion. 
3.7.2 Analysis of general data 
The basic demographics and information (such as age, gender and medical history, 
cancer stage, etc.) of the included cases are statistically described. The quantitative data 

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are expressed in forms of mean, standard deviation, median, minimum value, maximum 
value, lower quartile (Q1), upper quartile (Q3); categorical data are expressed in forms of 
the number of cases and the proportion. 
3.7.3 Statistical evaluation of test performance 
Evaluation indicators: positive percent agreement, negative percent agreement and 
overall percent agreement, and the corresponding 95% confidence interval. 
A four-fold table is used to calculate the positive percent agreement, negative percent 
agreement, positive predictive value, negative predictive value, overall percent agreement 
and 95% confidence interval of the test reagent and the control method. 
Table 1 Statistics of clinical results of test reagent and control method 
Test product 
Control method 
Total 
Positive 
Negative 
Positive 
A 
B 
A+B 
Negative 
C 
D 
C+D 
Total 
A+C 
B+D 
A+B+C+D 
Note: A sample with a mutation genotype is judged as positive, and a sample with no mutation 
genotype is judged as negative. 
The calculation formula of the percent agreement is as follows: 
Positive percent agreement =A/(A+C)×100%  
Negative percent agreement = D/(B+D) ×100%  
Overall percent agreement = (A+D)/(A+B+C+D)×100%  
The calculation formula of 95% confidence interval: p±1.96×[ p(1 -p)/n]1/2 (where p is 
the positive percent agreement, negative percent agreement, and overall percent agreement, 
n is the sample size. If p>0.9, use Wilson score method for correction). 
3.7.4 Statistical evaluation of diagnostic performance 
Table 2 Comparison of statistical results between the test reagent and clinical diagnostic standards 
Test product 
Clinical diagnostic criteria 
Total 
Positive 
Negative 
Positive 
A (true positive) 
B (false positive) 
A+B 

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Negative 
C (false negative) 
D (true negative) 
C+D 
Total 
A+C 
B+D 
A+B+C+D 
The calculation formula of the sensitivity is as follows: 
Sensitivity = A/(A+C)×100%  
Specificity = D/(B+D)×100%  
Overall percent agreement = (A+D)/(A+B+C+D)×100%  
Positive predictive value (PPV)=A/(A+B)×100%  
Negative predictive value (NPV)=D/(C+D)×100%  
The calculation formula of 95% confidence interval: p±1.96×[ p(1 -p)/n]1/2 (where p is 
sensitivity, specificity, positive predictive value, negative predictive value, total percent 
agreement, n is the sample size. If p>0.9, use Wilson score method for correction.) 
At the same time, compare the diagnostic performance of test reagent + AFP, test 
reagent + AFP + US, AFP + US to the clinical diagnostic criteria. 
3.7.5 Comparison of sensitivity across different test methods for screening liver cancer 
Make statistical tests on the sensitivity superiority of test reagent + AFP to 
conventional method (AFP+US) for screening liver cancer, the sensitivity superiority of 
test reagent + AFP + US to conventional method for screening liver cancer, the sensitivity 
non-inferiority of test reagent + AFP + US to conventional method for screening liver 
cancer, the sensitivity non-inferiority of test reagent + AFP + US to conventional method 
for screening liver cancer. 
3.8 Quality control of clinical trials 
Quality control is applied at every stage of clinical trials to ensure that all data are 
credible and correct. The quality control methods are as follows: 
1） Qualification of researchers participating in clinical trials: researchers 
participating in clinical trials must have professional expertise, qualifications and 
abilities in clinical trials, and pass the qualification review, and the personnel 
requirements should be relatively fixed. 
2） Training and pre-trial: The sponsor is responsible for the training of researchers 

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before the start of the clinical trial, so as to help the clinical researchers fully 
understand and understand the clinical trial protocol, the testing of clinical trial 
products, the filling of original records and forms, etc. know. 
3） Laboratory quality control: Clinical laboratories should strictly comply with 
relevant molecular biology laboratories, clinical The management of gene 
amplification laboratories is implemented. 
4） The test data is valid only if the control quality control test of the clinical trial 
product meets the requirements. 
5） Clinical trial monitoring: The monitor should formulate a complete monitoring 
plan before the start of the clinical trial, and monitor the clinical trial according to 
the monitoring plan to ensure that the clinical trial is carried out in accordance 
with the clinical protocol and ensure the original trial data. and completeness and 
accuracy of records. 
3.9 Controls for bias 
Subject samples should be blinded to avoid the introduction of bias due to the fact 
that the operator and the evaluator of the test results are aware of the subject's disease 
diagnosis or comparative reagent test results. The detection of in vitro diagnostic reagents 
for testing should be carried out simultaneously with the judgment of clinical reference 
standards or the detection of comparative reagents, so as to avoid the deviation of clinical 
trial conclusions from the true value due to different disease processes or large differences 
in sample storage time. Bias control is required in each stage of clinical trial institution 
selection, subject selection, trial process, and statistical analysis. 
3.10 
Data processing and record keeping 
Investigators should fill in the clinical trial case report form and record the contents 
of the form truthfully, in detail, and carefully to ensure that the contents in the form are 
complete, authentic and reliable. All observations and findings in the clinical trial should 
be verified to ensure the reliability of the data and to ensure that the conclusions in the 
clinical trial are derived from the original data. The clinical trial personnel should 
truthfully fill in the relevant clinical information and the test results of the clinical trial, 
and have a designated person to review and sign. 

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The clinical trial institution shall preserve the clinical trial data for 10 years after the 
end of the clinical trial. The sponsor shall save the clinical trial data until the medical 
device is not used.  
3.11 
Management of samples and instruments 
During the development of this trial, investigators who have been trained and 
qualified by the sponsor and authorized by the principal investigator shall independently 
manage the samples, reagents, and instruments used in the study to ensure the traceability 
of the preservation and use of the samples and reagents used in the study. in this test. 
3.12 
Auditing 
The sponsor should send qualified inspectors to monitor the trial process to ensure 
the quality of the trial. Inspectors should have corresponding professional backgrounds, 
have undergone necessary training, be familiar with relevant regulations and norms, be 
familiar with the non-clinical and clinical aspects of the in vitro diagnostic reagents for 
investigational and similar products, clinical trial protocols and related documents, and be 
familiar with the relevant documents. Follow the relevant standard operating procedures 
formulated by the sponsor, and urge the clinical trial to be implemented according to the 
protocol. 
4. Ethical issues and informed consent 
4.1 Ethical considerations 
This clinical trial strictly complies with relevant laws and regulations such as the 
"Quality Management Practice for Clinical Trials of Medical Devices" of the State Drug 
Administration and the "Declaration of Helsinki". This clinical validation trial involves 
patient-related data. Such as: patient's unique traceable number, age, gender, clinical 
diagnosis and imaging results, etc. The relevant information of the patient will be kept 
confidential by the hospital and Beijing Genetron Genetics Co., Ltd. The clinical trial 
implementer and clinical verification unit promise not to disclose the content related to the 
subject's specimen. 

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This clinical trial strictly follows the ethical guidelines established by the Declaration 
of Helsinki of the World Medical Association and the relevant regulatory requirements 
such as the "Measures for Ethical Review of Biomedical Research Involving Humans". 
This trial should be approved by the ethics committee of the clinical trial institution prior 
to initiation. Any modifications to the study protocol must also be notified and approved 
by the ethics committee before the changes are implemented. 
4.2 Informed consent process and informed consent text 
The informed consent form can only be used after it has been approved by the ethics 
committee of the clinical trial institution, and the revision of the informed consent form 
can only be implemented after the approval of the ethics committee of the clinical trial 
institution. 
1） Sign the informed consent form: 
The samples used in this clinical study are all prospective samples, which are 
specially collected for this clinical trial, and an informed consent form must be signed 
with the subjects. 
Before each subject is enrolled in this study, the research doctor is responsible for 
giving him or his designated representative a complete and comprehensive introduction to 
the purpose, procedures and possible risks of this study in writing. And inform the 
subjects to withdraw from the research at any time after notifying the investigator without 
discrimination or retaliation, and any medical treatment and rights will not be affected by 
this. 
2） Informed consent text 
The subjects screened and enrolled in this clinical trial are required to sign the 
informed consent. The sponsor provides the text of the informed consent form to the 
clinical trial institution, and obtains the consent and approval of the ethics committee of 
each clinical trial institution before the start of this clinical trial. 

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5. Analysis of the possibility of adverse events, and provisions for 
reporting adverse events and product defects 
This study will involve blood sample collection and examination with dynamic 
contrast-enhanced MRI. The collection of blood samples will be performed by 
professionals in strict accordance with aseptic requirements. There may be some very 
small risks in the collection of blood samples, including transient pain, local bruising, mild 
dizziness in a few people, or extremely rare needle infection; The injection of contrast 
agent used in dynamic contrast-enhanced MRI examination may cause the risk of allergy 
to the human body; if some subjects are claustrophobic, it may lead to disease attacks, and 
claustrophobia should be inquired in advance and excluded when subjects are screened 
patient. If there is any damage related to the blood sample collection and the contrast 
agent used in the dynamic enhanced MRI in this clinical study, the investigator will treat 
the subject according to the specific situation of the subject to ensure the safety of the 
subject and the cost of the treatment. shall be borne by the sponsor. 
5.1 Adverse event 
Adverse events refer to adverse medical events that occur during clinical trials of 
medical devices, regardless of whether they are related to the experimental medical 
devices. 
In the event of adverse events related to this study, the principal investigator should 
provide the subjects with adequate and timely treatment and treatment, and inform the 
subjects in a timely manner when the subjects develop concurrent diseases that require 
treatment and treatment. 
5.2 Serious adverse event 
Serious adverse events refer to medical device clinical trials that lead to death or 
serious deterioration of health status, including fatal diseases or injuries, permanent 
defects in body structure or function, requiring hospitalization or prolonged hospitalization, 

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and requiring medical treatment. To avoid permanent defects in body structure or function; 
events such as fetal distress, fetal death, or congenital anomalies and congenital defects. 
The principal investigator should report to the sponsor, the management department 
of the medical device clinical trial institution, and the ethics committee within 24 hours 
after the serious adverse event is known, to ensure that the subjects receive appropriate 
treatment and follow-up, and submit a follow-up report and/or summary of serious adverse 
events Report. When necessary, report to the drug regulatory department and health 
management department of the province, autonomous region, or municipality directly 
under the Central Government where it is located. 
After the first serious adverse event report, the investigator should actively follow up 
the patient and provide the sponsor or designee with further information about the patient's 
condition and follow up until the event resolves, the condition has stabilized, the event is 
otherwise reasonably explained or accepted The subjects were lost to follow-up. 
The sponsor or designee may request or schedule additional examinations and/or 
evaluations by the investigator to clarify the nature and/or causality of the serious adverse 
event to the extent possible. It is the responsibility of the investigator to provide support. If 
a patient dies while participating in the study, a copy of any autopsy report should be 
provided to the sponsor. 
5.3 Device defect 
Device defects refer to the unreasonable risks of medical devices that may endanger 
human health and life safety under normal use conditions during clinical trials, such as 
label errors, quality problems, malfunctions, etc. 
Some of the reagents in this clinical trial are stored at low temperature, and this part 
of the reagents may fail the sample test due to improper storage. During the whole 
experimental process, the reagents and samples involved in this experiment were stored in 
accordance with the corresponding instructions and the requirements of this clinical trial 
protocol. In the event of the above situations, the investigators should timely and truthfully 
record them in the relevant trial records, and analyze and summarize them in the clinical 

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trial report. 
6. Provisions on the revision of clinical trial protocols 
After the trial protocol is determined and approved by the ethics committee, its 
clinical research design should not be changed in general, but may be modified in the 
following circumstances: During the trial process, if it is found that it is difficult to select 
qualified samples according to the original inclusion/exclusion criteria , it is necessary to 
analyze the reasons and take corresponding measures to modify the original 
inclusion/exclusion criteria. 
Any modification to the trial protocol should be stated in the revised protocol, which 
must be agreed by the sponsor, the clinical trial institution and the investigator, and the 
reasons for the modification must be recorded. The revised protocol needs to be 
re-approved by the ethics committee. 
7. What should be covered in a clinical trial report 
After the clinical trial is over, the investigators of each clinical trial institution shall 
summarize their respective clinical trial data, issue a clinical trial summary, and attach the 
clinical trial data sheet, other test methods used in the clinical trial, or other basic 
information on in vitro diagnostic reagents and other products. and other information. 
The clinical trial report should include the names, units, responsibilities in the 
research, clinical trial methods, enrolled cases, sample allocation, result analysis, etc. 
8. Confidentiality 
The personal data of the subjects participating in this clinical study in the trial are 
kept confidential. The research physician and other researchers will use the subject's 
medical information to conduct the research. This information may include the subject's 
name, address, telephone number, medical history, and information obtained during your 
study visit. Subject files are to be kept in locked filing cabinets for researcher access only. 
A number is used in the study to identify the subject's research information and laboratory 
test specimens. Only investigators and research team members can look up the number. In 

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order to ensure that the research is carried out in accordance with the regulations, when 
necessary, the research sponsor, the members of the government management department 
or the ethics review committee can access the personal data of the subjects in the research 
unit according to regulations. When the results of this study are published, no personal 
information on the subjects will be disclosed. 
9. Responsibilities of each party 
9.1 Responsibilities of the sponsor 
1) Provide free test products that have passed the inspection, and provide test-related 
documents, research funds, etc. 
2) As the responsible party for quality assurance of clinical trials, qualified inspectors 
should be dispatched to monitor the quality of the trials to ensure that all trial data meet 
relevant requirements. 
3) Select a qualified clinical trial institution to jointly study and formulate a clinical trial 
plan with the principal investigator. 
4) Responsible for training the researchers of the clinical trial institution related to the 
trial. 
5) The institution and the principal investigator should be informed of problems in the trial 
in a timely manner, so that the institution can take relevant measures to improve and 
protect the subjects. 
6) Before deciding to suspend or suspend a clinical trial, the research institution, 
investigator and ethics committee must be notified in writing, and the reasons shall be 
stated. 
7) After the trial, submit the final report and clinical trial summary (sub-center) report to 
the Ethics Committee and Institutional Office as required. 
8) Without the subject's written consent, the subject's personal information/specimens 
cannot be used for commercial purposes or other research without authorization. 
9.2 Responsibilities of clinical trial institutions 
1) Design and formulate a clinical trial protocol together with the sponsor, and strictly 

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follow the protocol. 
2) Ensure that the data are truthfully, accurate, complete, timely and lawful in the medical 
records and case report forms. 
3) In case of serious adverse events caused by the assessment products, appropriate 
treatment measures should be taken immediately for the subjects, and at the same time, 
they should be reported to the institutional ethics committee and the sponsor, and reported 
to the relevant departments within the specified time. 
4) Accept the supervision of the supervisor dispatched by the sponsor and the inspection 
of the drug regulatory department to ensure the quality of the clinical trial. 
5) The information and clinical trial data provided by the sponsor shall not be leaked 
without the consent of the sponsor. 
6) If the trial progress is affected or the trial is interrupted due to the quality problem of 
the trial product and the failure to provide the trial product and consumables in time, the 
clinical trial institution shall not be held responsible. 
7) Responsible for writing the clinical trial summary (summary) report, signed and dated 
by the clinical trial personnel, and handed over to the sponsor after the clinical trial 
institution seals it.