针对具有遗传性癌症倾向的个体的个性化监测、预防和癌症治疗选择取决于种系基因检测的结果。基因组技术的改进，例如 RNA 测序的可用性，可以通过提高种系检测的准确性和产量来增加对有资格进行个性化干预的个体的识别。评估配对 DNA 和 RNA 检测与致病种系检测的累积关联遗传变异和不确定意义变异的解析 (VUS)。2019 年 3 月至 2020 年 4 月，对在单一诊断实验室接受遗传性癌症指征种系检测的个体进行了配对 DNA 和 RNA 测序。人口统计特征、临床数据和检测结果在收到样本时进行整理，并随着时间的推移评估变体分类的变化。数据分析于2020年5月至2023年6月进行。主要结果是诊断率提高、VUS率降低、变异类型的总体结果、RNA证据与变异分类的关联以及对癌症风险管理的相应预测效果。总共纳入了 43524 名个体（测试时的中位年龄 [范围]，54 [2-101] 岁；37373 名女性个体 [85.7%]，6224 名男性个体 [14.3%]，以及 2 名性别未知的个体 [< 0.1%]），进行了 43599 次测试。共有 2197 名（5.0%）德系犹太人，1539 名（3.5%）亚裔，3077 名（7.1%）黑人，2437 名（5.6%）西班牙裔，27793 名（63.7%）白人，2049 名（4.7%） ）是其他种族，其中 4507 人（10.3%）的种族和民族未知。 549 人 (1.3%) 的变异分类受到影响。 97 名个体进行了具有医学意义的升级，其中 70 名个体的变异从 VUS 重新分类为致病性/可能致病性 (P/LP)，27 名个体具有新的深部内含子 P/LP 变异，而使用 DNA 无法检测到该变异单独测序。 545 个 P/LP 剪接变异中共有 93 个 (17.1%) 依​​赖于 RNA 证据进行分类，439 个现有剪接 VUS 中的 312 个 (71.1%) 通过 RNA 证据解决。值得注意的是，与白人相比，亚洲人、黑人和西班牙裔人的阳性率上升 (3.1%) 和 VUS 率下降 (-3.9%) 更高（1.6%；P = .02；和 -2.5%； P < .001)。这项诊断研究的结果表明，同时进行 RNA 测序和 DNA 测序的能力通过改进新变异的检测和现有变异的分类，代表了种系基因检测的重要进步。这扩大了对具有遗传性癌症倾向的个体的识别，并增加了个性化治疗和监测的机会。

Personalized surveillance, prophylaxis, and cancer treatment options for individuals with hereditary cancer predisposition are informed by results of germline genetic testing. Improvements to genomic technology, such as the availability of RNA sequencing, may increase identification of individuals eligible for personalized interventions by improving the accuracy and yield of germline testing.To assess the cumulative association of paired DNA and RNA testing with detection of disease-causing germline genetic variants and resolution of variants of uncertain significance (VUS).Paired DNA and RNA sequencing was performed on individuals undergoing germline testing for hereditary cancer indication at a single diagnostic laboratory from March 2019 through April 2020. Demographic characteristics, clinical data, and test results were curated as samples were received, and changes to variant classification were assessed over time. Data analysis was performed from May 2020 to June 2023.Main outcomes were increase in diagnostic yield, decrease in VUS rate, the overall results by variant type, the association of RNA evidence with variant classification, and the corresponding predicted effect on cancer risk management.A total of 43 524 individuals were included (median [range] age at testing, 54 [2-101] years; 37 373 female individuals [85.7%], 6224 male individuals [14.3%], and 2 individuals of unknown sex [<0.1%]), with 43 599 tests. A total of 2197 (5.0%) were Ashkenazi Jewish, 1539 (3.5%) were Asian, 3077 (7.1%) were Black, 2437 (5.6%) were Hispanic, 27 793 (63.7%) were White, and 2049 (4.7%) were other race, and for 4507 individuals (10.3%), race and ethnicity were unknown. Variant classification was impacted in 549 individuals (1.3%). Medically significant upgrades were made in 97 individuals, including 70 individuals who had a variant reclassified from VUS to pathogenic/likely pathogenic (P/LP) and 27 individuals who had a novel deep intronic P/LP variant that would not have been detected using DNA sequencing alone. A total of 93 of 545 P/LP splicing variants (17.1%) were dependent on RNA evidence for classification, and 312 of 439 existing splicing VUS (71.1%) were resolved by RNA evidence. Notably, the increase in positive rate (3.1%) and decrease in VUS rate (-3.9%) was higher in Asian, Black, and Hispanic individuals combined compared to White individuals (1.6%; P = .02; and -2.5%; P < .001).Findings of this diagnostic study demonstrate that the ability to perform RNA sequencing concurrently with DNA sequencing represents an important advancement in germline genetic testing by improving detection of novel variants and classification of existing variants. This expands the identification of individuals with hereditary cancer predisposition and increases opportunities for personalization of therapeutics and surveillance.