BD Rhapsody Onco-BC 乳癌靶向檢測試劑盒（panel）

英文名稱：BD Rhapsody™  Onco-BC Targeted Panel

中文名稱：BD Rhapsody Onco-BC 乳癌靶向檢測試劑盒（panel）

用途：與BD Rhapsody單細胞表達分析儀配合使用，檢測與乳腺癌和免疫細胞相關的 400 個基因

乳腺癌生物學因復雜的基因表達，細胞異質(zhì)性和患者間的差異而變得復雜。為了解決這一復雜性，BD Rhapsody™OncoBC靶向檢測panel使用BD Rhapsody單細胞分析系統(tǒng)為單細胞乳腺癌研究提供了一種簡化的檢測方法。 通過集中分析與乳腺癌生物學zui相關的基因和標志物，減少測序通量，并且降低了與整個轉(zhuǎn)錄組測序相關時間和測序成本。

BD Rhapsody™  Onco-BC Targeted Panel特點

經(jīng)驗證的基因panel專注于人類乳腺癌生物學

有針對性的方法提高了實驗效率

可重復的檢測方法可用于各種實驗

歡迎您咨詢華雅再生醫(yī)學旗艦公司：紅榮微再（上海）生物工程技術有限公司  ：1500 1904 520。紅榮微再-客服:  經(jīng)銷商專員

BD Rhapsody單細胞分析系統(tǒng)

BD Rhapsody單細胞分析系統(tǒng)是一種新型的單細胞mRNA表達分析儀器。它能捕獲并分離細胞，通過磁珠對細胞和mRNA轉(zhuǎn)錄本進行分類擴增以供測序。作為單細胞RNA-seq的新方法，與傳統(tǒng)的全轉(zhuǎn)錄組擴增（WTA）相比，BD Rhapsody單細胞分析系統(tǒng)可以同時處理12個樣本，能夠?qū)Τ汕先f個單細胞的數(shù)百個表達基因進行數(shù)字定量，極大程度節(jié)省實驗時間和降低測序成本。

BD Rhapsody單細胞分析系統(tǒng)可以定制靶向組合（panel），靶向方法有助于提高單細胞測序效率，并推動對當今一些挑戰(zhàn)性的科學領域的進一步探索。BD經(jīng)驗證的檢測試劑盒（panel）顯示穩(wěn)定且可重現(xiàn)的細胞異質(zhì)性陣列，可用于探索與眾多應用相關的細胞類型。BD Rhapsody 系統(tǒng)包括以下檢測試劑盒（panel）：

□ BD Rhapsody Onco-BC 靶向檢測試劑盒（panel） – 與乳腺癌和免疫細胞相關的 400 個基因

□ BD Rhapsody 免疫反應檢測試劑盒（panel） – 免疫細胞類型譜分析

□ BD Rhapsody  T-細胞靶向檢測試劑盒（panel）

□ BD Rhapsody 補充檢測試劑盒（panel）

□ BD Rhapsody 自定義檢測試劑盒（panel）

BD Rhapsody Onco-BC 靶向檢測試劑盒（panel）

Panel designed by scientists, for scientists

The customizable and validated BD Rhapsody Onco-BC Targeted Panel was created from collaborations with clinical oncologists.

The panel content combines data from experimental and database sources, chosen to include genes with known involvement in cancer progression, proliferation, epithelial-tomesenchymal transition (EMT) and cancer stem cells. Including PAM50 genes aids molecular subtyping of breast cancer cells¹,while immunophenotyping and stromal cell markers enable comprehensive surveys of the complex tumor microenvironment.

科學家為科學家設計的Panel

可定制且經(jīng)過驗證的BD Rhapsody Onco-BC乳腺癌靶向Panel是由臨床腫瘤學家合作開發(fā)的。

BD Rhapsody Onco-BC乳腺癌靶向Panel結合了來自實驗和數(shù)據(jù)庫來源的數(shù)據(jù)，選擇的數(shù)據(jù)包括已知參與癌癥progression，擴增，上皮 - 間質(zhì)轉(zhuǎn)化（EMT）和癌癥干細胞的基因。 包括PAM50基因有助于乳腺癌細胞的分子亞型¹，而免疫分型和基質(zhì)細胞標志物可以對復雜的腫瘤微環(huán)境進行全面調(diào)查。

Tailored libraries for targeted sequencing

The BD Rhapsody Onco-BC Targeted Panel is designed for the BD Rhapsody system, and uses multiplex PCR for detecting up to 389 genes. After cells are lysed in the BD Rhapsody Cartridge,

beads hybridized with mRNA are magnetically retrieved for cDNA synthesis. Primers included in the BD Rhapsody Onco-BC Targeted Panel and BD Rhapsody Supplemental Panel are used for several rounds of gene-specific, nested PCR for library construction. Final PCR products for sequencing contain adapters, a cell label, a unique molecular index (UMI), and up to 400 bp of the 3’ end of a target gene. Assay products may be sequenced on Illumina® MiniSeq, MiSeq, NextSeq, HiSeq2500 and HiSeq4000 sequencers.

靶向測序的定制文庫

BD Rhapsody Onco-BC乳癌靶向Panel專為BD Rhapsody系統(tǒng)設計，使用多重PCR檢測多達389個基因。細胞在BD Rhapsody Cartridge中裂解后，與mRNA雜交的磁珠被磁性回收用于cDNA合成。包含在BD Rhapsody Onco-BC靶向Panel和BD Rhapsody Supplemental Panel 中的引物用于多輪基因特異性巢式PCR，用于構建文庫。用于測序的zui終PCR產(chǎn)物含有接頭，細胞標記（label），*的分子索引（unique molecular index，UMI）和3'末端高達400bp的靶基因。檢測產(chǎn)物可以在Illumina®MiniSeq，MiSeq，NextSeq，HiSeq2500和HiSeq4000測序儀上進行測序。

Figure 1. cDNA synthesis and library construction chemistry with the BD Rhapsody Onco-BC Targeted Panel.

圖1.BD Rhapsody Onco-BC Targeted Panel的cDNA合成和文庫構建原理

Your target genes, delivered your way

Researchers may select up to 100 additional genes for analysis,with the BD Rhapsody Supplemental Panel. Using BD’s primer design software, primers for supplemental gene targets are designed to be compatible with pre-designed base panels.

Supplemental primers are shipped to you and combined with the BD Rhapsody Onco-BC Targeted Panel for library preparation.

專屬目標基因定制

研究人員可以經(jīng)由BD Rhapsody Supplemental Panel選擇zui多100個額外基因分析。使用BD的引物設計軟件，補充目標基因的引物可以與預設基礎panel兼容。

Remarkable panel performance—single-cell suspensions from a metastatic breast cancer sample

In the following example, cryopreserved single-cell suspensions from a disaggregated metastatic breast cancer patient sample were sorted using the BD FACSAria™ system to enrich for live cells. 4,409 cells derived from a lymph node metastasis were profiled using the BD Rhapsody system. Fifteen cell types were detected, including four subpopulations of breast cancer cells and a diverse tumor microenvironment of stromal and major immune cell types. Because the analysis focused on just five percent of the transcriptome, the amount of sequencing is drastically lowered.

的panel性能—轉(zhuǎn)移性乳腺癌樣本的單細胞懸液

在下面的實例中，使用BD FACSAria ^TM系統(tǒng)分選來自disaggregated轉(zhuǎn)移性乳腺癌患者樣品的凍存單細胞懸浮液以富集活細胞。使用BD Rhapsody系統(tǒng)對來源于淋巴結轉(zhuǎn)移的4,409個細胞進行分析。檢測到15種細胞類型，包括4種乳腺癌細胞亞群和多種腫瘤微環(huán)境（間質(zhì)細胞和主要免疫細胞組成）。由于分析只集中在轉(zhuǎn)錄組的5％，因此測序通量極大降低。

Table 1: Example genes by category

Pathway # of genes in panel

Cell adhesion 23；Cell cycle 44；Chemokine/chemokine receptor,cytokine/cytokine receptor 53；

Signal transduction 56；Oncogenes, tumor suppressors 10；Tumor immunity, immune checkpoint,

immune cell activation 24；Surface antigen 32；Other* 147（ Angiogenesis; cell death; complement system; structural; transport; DNA repair;epithelial-to-mesenchymal transition; hormone receptor; immunoglobulin; lncRNA;metabolism; multi-drug resistance; protease; transcription factor）

表1 參考基因

其他：血管生成; 細胞死亡; 補充體系;結構; 運輸; DNA修復;上皮間質(zhì)轉(zhuǎn)化; 激素受體;免疫球蛋白;lncRNA;代謝; 多重耐藥性;蛋白酶; 轉(zhuǎn)錄因子

表2：BD Rhapsody Onco-BC Targeted Panel metrics

BD Rhapsody Onco-BC乳癌靶向檢測panel應用實例

轉(zhuǎn)移性乳腺癌樣本分析

Analysis of a metastatic breast cancer sample

Figure 2. Example data from analysis of a lymph node containing metastatic BrCa cells using the BD Rhapsody Onco-BC Targeted Panel. (a) tSNE projection of 4,409 cells annotated by cell type and (b) box plots of marker gene expression for each major cell type show complex tumor heterogeneity.

圖2.使用BD Rhapsody Onco-BC靶向Panel分析含有轉(zhuǎn)移性BrCa細胞淋巴結的實例數(shù)據(jù)。 （a）細胞類型注釋的4,409個細胞的tSNE projection和（b）每種主要細胞類型的標志基因box plots顯示復雜的腫瘤異質(zhì)性。

Learn more about the power of targeted single-cell analysis

The BD Rhapsody Onco-BC Targeted Panel brings single-cell analysis within reach of all breast cancer researchers, by combining increased sequencing efficiency with a flexible approach to your projects. Contact us to find out how the BD Rhapsody panel can help you further the understanding of breast cancer biology.

了解更多單細胞靶向分析功能

BD Rhapsody Onco-BC Targeted Panel通過將更高的測序效率與靈活的方法結合到您的項目中，使所有乳腺癌研究人員都能夠進行單細胞分析。，了解BD Rhapsodypanel如何幫助您進一步了解乳腺癌生物學。

參考文獻

1. Parker JS, Mullins M, Cheang MCU, et al. Supervised Risk Predictor of Breast Cancer Based on Intrinsic Subtypes. Journal of Clinical Oncology. 2009;27(8):1160-1167.

歡迎您咨詢華雅再生醫(yī)學旗艦公司：紅榮微再（上海）生物工程技術有限公司  ：1500 1904 520。紅榮微再-客服:  經(jīng)銷商專員