Oncomine Lung 无细胞总核酸研究检测
Oncomine Lung 无细胞总核酸研究检测
Ion Torrent™

Oncomine Lung 无细胞总核酸研究检测

Oncomine Lung cfTNA 研究检测是检测从全血血浆组分中分离的肺癌肿瘤来源无细胞 DNA 和 RNA(无细胞总核酸;cfTNA)的完整解决方案的一部分。其提供了文库构建试剂,以及用于从单管了解更多信息
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货号数量
A358648 reactions
货号 A35864
价格(CNY)
-
数量:
8 reactions
Oncomine Lung cfTNA 研究检测是检测从全血血浆组分中分离的肺癌肿瘤来源无细胞 DNA 和 RNA(无细胞总核酸;cfTNA)的完整解决方案的一部分。其提供了文库构建试剂,以及用于从单管 10 mL 全血的血浆组分中获得的 cfTNA 制备扩增子文库的一池多重 PCR 引物。

液体活检相对于传统实体瘤活检具有几大优势:
•侵入性较小,可在多个时间点采集样品以监测癌症进展
•成本较低
•从样品到结果的周转时间更短
•更好地表示肿瘤异质性

Oncomine Lung cfTNA 研究检测可实现以下分析:
• 热点基因 (SNV) 和短插入缺失:ALK、BRAF、EGFR、ERBB2、KRAS、MAP2K1、MET、NRAS、PIK3CA、ROS1 和 TP53(覆盖 ∼168 个热点)
• 基因融合:ALK、RET、ROS1
• MET 外显子 14 跳读
• 拷贝数基因 (CNV):MET

已确定这些基因在非小细胞肺癌 (NSCLC) 中经常突变。通过使用标签测序技术,可以实现各种变体类型的检测下限 (LOD)*:
•对于 SNV/短插入缺失,可实现 LOD 为 0.1%、灵敏度 ∼90%、特异性 >99%
• 对于融合和 MET 外显子跳读,可实现 LOD 为 1%、灵敏度 >90%、特异性 >99%
• 对于 MET CNV 靶标,可实现检测低至 1.2 倍扩增、灵敏度 >90%、特异性 >99%

使用 Ion S5 XL 测序系统可仅在 2 天内完成从使用 MagMAX 无细胞总核酸分离试剂盒进行 cfTNA 分离到样品分析的整个工作流程。

技术
在全血血浆组分中观察到极低浓度 cfDNA 和 cfRNA。由于此低发病率,所以该检测中采用了标签测序技术。该技术将独特的分子标签连接至基因特异性引物上(下图)。扩增后,将根据标签对已标记分子进行分组。在 80% 或更长的时间含有相同突变体的组将被称为阳性。使用标签技术,通过文库构建/测序过程生成的随机错误序列能够被清除。

与 LOD 为 1-5% 的其他技术不同,Oncomine 肺 cfTNA 研究测定试剂盒针对 SNV 具有低至 0.1% 的灵活检测限或在 1,000 个野生型拷贝背景中具有 1 个突变体拷贝。为了达到 0.1% LOD,需要 20 ng cfDNA 上样量。cfTNA 用量可以更低,但 LOD 将更高,这取决于起始量。

优势:
•用于短 cfDNA 的优化扩增子设计确保可能较高的捕获率
• 标签测序技术通过去除随机掺入的错误,尽可能减少假阳性结果
• 优化的靶向测定试剂盒设计可进行高度多重下一代测序 (NGS),
降低每份样品的测序成本
• 2 天内即可完成的高效工作流程

使用 Torrent Suite 软件 5.2 或更高版本,可以实现对 SNV 和短插入缺失的分析。为分析 SNV、短插入缺失、融合和 CNV,需要 Ion Reporter 5.6(基于云端或服务器)。

标签测序技术具有简单性、快速和可扩展性
Oncomine 肺 cfTNA 研究测定试剂盒只需 5 ng cfTNA 上样量即可进行靶向文库构建从而进行癌症基因研究。Oncomine Lung cfTNA 研究检测与 FFPE 样品兼容,可用于潜在的一致性研究。获得靶向文库的总时间仅为 4 小时。使用标签测序条形码套件 1-24 或 25-48(货号 A31830、A31847)实现可扩展性和灵活性,用于在 Ion S5 芯片上倍增条形码样品。

*使用从正常健康供体中分离的人工阳性样品和 cfTNA,测定各变体类型的敏感性和特异性。
仅供科研使用。不可用于诊断程序。
规格
适用于(应用)测序
反应次数8
产品线Oncomine™
产品类型Oncomine Lung 无细胞总核酸研究检测
数量8 reactions
运输条件干冰
Unit SizeEach
内容与储存
• 16 μL lung cfTNA 检测板
• 320 μL cfDNA 文库 PCR 预混液
• 832 μL 低 TE 缓冲液
• 8 μL cfDNA 文库引物 P1
• 8 μL cfDNA 文库引物 A/BC1
• 22 μL SuperScript VILO 预混液

常见问题解答 (FAQ)

For the Oncomine Lung Cell-Free Total Nucleic Acid Research Assay, what are the analysis workflows in Ion Reporter Software?

There are two analysis workflows available:

1. Oncomine TagSeq Lung v2 Liquid Biopsy - w2.0 - Single Sample: Detects and annotates low-frequency variants including SNPs/Indels (down to 0.1% limit of detection), Fusions, and CNVs from targeted nucleic acid libraries (DNA & RNA) from the Oncomine Lung Cell‑Free Total Nucleic Acid Research Assay. This is compatible with DNA & RNA purified from cell-free total nucleic acids.

2. Oncomine TagSeq Lung v2 Tumor - w2.0 - Single Sample: Detects and annotates low-frequency variants including SNPs/Indels (down to 0.5% limit of detection), fusions, and CNVs from targeted nucleic acid libraries (DNA & RNA) from the Oncomine Lung Cell-Free Total Nucleic Acid Research Assay. Due to deamination events caused by the FFPE process, the minimum alternative allele frequency is set to 0.3%. This makes it compatible with DNA & RNA purified from FFPE tumor tissue as well as fresh frozen tumor tissue.

Are there any BED files for the Oncomine Lung Cell-Free Total Nucleic Acid Research Assay?

Yes, there is a specific BED file and Hotspot file for the Oncomine Lung Cell-Free Total Nucleic Acid Research Assay. Please contact your local Field Application Specialist (FAS) or Clinical Application Consultant (CAC) to request the BED files.

What is the LOD (limit of detection) for the Oncomine Lung Cell-Free Total Nucleic Acid (cfNA) Research Assay?

Through the use of Tag Sequencing technology, low limits of detection (LOD) can be achieved for different variant types*:

- For SNVs/short indels, an LOD of 0.1% can be achieved with sensitivity of ˜90% and specificity of >99%
- For fusions & MET exon skipping, an LOD of 1% can be achieved with sensitivity of >90% and specificity of >99%
- For MET CNV target, detection as low as 1.2-fold amplification can be achieved with sensitivity of >90% and specificity of >99%

*Sensitivity and specificity for each variant type were determined using a collection of contrived positive samples and cfNA isolated from normal healthy donors.

Does the Oncomine Lung Cell-Free Total Nucleic Acid Research Assay use AmpliSeq technology?

No, this assay uses Tag Sequencing technology. Cell-free DNA (cfDNA) and cell-free RNA (cfRNA) are found at extremely low concentrations in the plasma fraction of whole blood. Because of this low prevalence, Taq Sequencing technology is utilized in this assay. The technology attaches unique molecular tags to the gene-specific primers. After amplification, the tagged molecules are grouped based on the tags. Groups containing the same mutant variant 80% of the time or greater will be called positive. Using the Tag technology, groups that contain random errors generated through the library construction/sequencing process are removed.

Which kit should I use to isolate cell-free total nucleic acid (cfNA) for the Oncomine Lung Cell-Free Total Nucleic Acid Research Assay?

Use cell-free total nucleic acid (cfNA) extracted using a method optimized for cfNA isolation from plasma. We recommend the MagMAX Cell Free Total Nucleic Acid Isolation Kit (Cat. No. A36716). You can expect 5-50 ng of cfDNA and 5-100 pg of cfRNA from 10 mL blood research sample collected in a K2 EDTA blood collection tube (Cat. No. ??)