Clinical Application of Liquid Biopsy for Precise Diagnosis and Prognosis in Lymphoma
Clinical Application of Liquid Biopsy for Precise Diagnosis and Prognosis in Lymphoma
Lymphoma is a highly heterogeneous blood malignancy. It is very important to search for
 relative specific diagnostic markers that can detect related lymphoma in early stage for the
 treatment and long-term prognosis of the disease, as the hematopoietic diseases, such as
 lymphoma, are more difficult to biopsy than solid tumors, with more damage and side
 effects.Liquid Biopsy (Liquid Biopsy) refers to the extraction of solid biological tissue, is
 the most common blood, also including saliva, urine, cerebrospinal fluid and other body
 fluids, and extract the circulating tumor cells (circulating tumor cell, CTC) and circulating
 tumor DNA (circulating tumor DNA, ctDNA) is used to assess related diseases. CTCs/CSCs have
 the ability to generate new tumors and play a key role in tumor metastasis.This project
 intends to develop liquid biopsy technology for accurate diagnosis and prognosis judgment of
 lymphoma, to carry out clinical transformation application and serve patients.
1. Patients with series and matched lymphoma who were refractory to initial diagnosis,
 remission and recurrence were enrolled, our research group will collected patient
 information comprehensively and signed the informed consent.

 2. The standard diagnosis process and treatment selection will be completed, and our
 research group willcollect relevant clinical and laboratory data.

 3. 5 ml of peripheral blood from each patient was collected with EDTA anticoagulant tube,
 and the blood cells were centrifuged at 820×g for 10 min at 4℃ within 1 h to separate
 the blood cells.The plasma was then transferred to a microcentrifuge tube and
 centrifuged at 4℃ at 20,000×g for 10 min to remove cell debris.According to the reagent
 instructions, cfDNA was extracted from 2 ml cell-free plasma and the concentration of
 cfDNA in each patient was determined.Meanwhile, genomic DNA kit was used to extract
 matching tumor DNA from lymphoma bone marrow tissue.

 4. We will extract the lymphoma of bone marrow tissue samples of RNA, and reverse
 transcription for cDNA, the use of qRT PCR verifying the mutations in tissue samples,
 after identifying the lymphoma five of the most common mutations from the cancer genome
 map (TCGA;http://cancergenome.nih.gov/) .

 5. The differences of mutations detected by liquid biopsy and tumor in situ biopsy will be
 compared, by NGS sequencing of peripheral blood cfDNA and lymphoma tissue DNA.
 Meanwhile, the mutation rate of each gene mutation was analyzed for lymphoma tissue
 type.

 6. The classification and malignant changes of matched lymphomas were statistically
 analyzed to study the differences in cfDNA concentration of different types of
 lymphomas, according to the cfDNA concentration determined at the earlier stage.

 7. The mutation differences of cfDNA in the peripheral blood of each patient will be
 compared at three time points before, during and after treatment, combined with PET/CT
 detection, and the correlation between cfDNA detection indexes and therapeutic effect
 and prognosis monitoring will be evaluated.

 8. SPSS 23.0 statistical software would be used to evaluate the correlation between cfDNA
 concentration in lymphoma and disease staging, grouping, subtype, efficacy prediction
 and recurrence monitoring. P<0.05 was considered statistically significant.
Inclusion Criteria:

 -  patients of lymphoma diagnosed of de novo, CR, relapsed/refractory, be willing to
 receive treatment

 Exclusion Criteria:

 -  leukemia