呼气凝析液中miRNA检测的新一代测序:一项初步研究。

IF 3.2 Q2 GENETICS & HEREDITY
Roberto Cherchi, Roberto Cusano, Sandro Orrù, Paolo A Ferrari, Matteo Massidda, Giorgio Fotia, Sara De Matteis, Pierluigi Cocco
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引用次数: 0

摘要

呼气冷凝物(EBC)取样已被认为是一种侵入性低、成本效益高的生物大分子检测方法,包括miRNA。为了探索其作为石棉暴露早期影响的生物标志物的可行性,我们对男性志愿者进行了初步测试,使用两种不同的测序平台比较了EBC和血浆中的miRNA谱。方法:六名男性志愿者参加了测试,他们都是退休的,没有接触过灰尘或烟雾。从200 μL的EBC样本和相同大小的血浆样本中提取RNA。样品等分在2个实验室使用2种不同的测序平台进行处理:MiSeq Illumina®平台和性能更高的HiSeq Illumina®平台。结果:HiSeq3000®测序平台鉴定出的独特分子指数(UMI)验证的miRNA数量是MiSeq®平台的两倍。在两种平台中,5/6名受试者的EBC计数与血浆计数的Spearman相关系数均显著(MiSeq®= 0.128-0.508,P = 0.026 -®= 0.156-0.412,P =。001-®平台显著提高了EBC miRNA的检出率。结论:我们的初步研究证实,当使用HiSeq®测序平台时,EBC取样是一种适合的、无创的方法来检测健康受试者的miRNA谱。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Next Generation Sequencing for miRNA Detection on the Exhaled Breath Condensate: A Pilot Study.

Next Generation Sequencing for miRNA Detection on the Exhaled Breath Condensate: A Pilot Study.

Next Generation Sequencing for miRNA Detection on the Exhaled Breath Condensate: A Pilot Study.

Next Generation Sequencing for miRNA Detection on the Exhaled Breath Condensate: A Pilot Study.

Introduction: Exhaled breath condensate (EBC) sampling has been suggested as a less-invasive and cost-effective method to detect biological macromolecules, including miRNA. To explore the feasibility of its use as a biomarker of early effects of asbestos exposure, we conducted a preliminary test on male volunteers by comparing the miRNA profile in the EBC and the plasma using 2 different sequencing platforms.

Methods: Six male volunteers, all retired and unexposed to dust or fumes, participated in the test. RNA was extracted from 200 μL EBC samples and same-size plasma samples. Sample aliquots were processed in 2 laboratories using 2 different sequencing platforms: a MiSeq Illumina® platform and a more performing HiSeq Illumina® platform.

Results: The HiSeq3000® sequencing platform identified twice as many unique molecular indexes (UMI)-validated miRNA as the MiSeq® platform. The Spearman's correlation coefficient between EBC counts and plasma counts was significant in 5/6 subjects with either platform (MiSeq® = 0.128-0.508, P = .026-<.001; HiSeq® = 0.156-0.412, P = .001-<.001). The intraclass correlation coefficient confirmed the consistency of the miRNA profile over the 6 participants with both biospecimens. Exploring the agreement between the EBC and plasma samples with Bland-Altman plots showed that using the HiSeq3000® platform substantially improved the EBC miRNA detection rate.

Conclusion: Our preliminary study confirms that, when using the HiSeq® sequencing platform, EBC sampling is a suitable, non-invasive method to detect the miRNA profile in healthy subjects.

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来源期刊
Epigenetics Insights
Epigenetics Insights GENETICS & HEREDITY-
CiteScore
5.10
自引率
0.00%
发文量
10
审稿时长
8 weeks
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