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Single Particle Inductively Coupled Plasma Mass Spectrometry-Based Homogeneous Detection of HBV DNA with Rolling Circle Amplification-Induced Gold Nanoparticle Agglomeration
Analytical Chemistry ( IF 6.7 ) Pub Date : 2022-07-05 , DOI: 10.1021/acs.analchem.2c00272 Yan Xu 1 , Guangyang Xiao 1 , Beibei Chen 1 , Man He 1 , Bin Hu 1
Analytical Chemistry ( IF 6.7 ) Pub Date : 2022-07-05 , DOI: 10.1021/acs.analchem.2c00272 Yan Xu 1 , Guangyang Xiao 1 , Beibei Chen 1 , Man He 1 , Bin Hu 1
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A highly sensitive and simple method based on rolling circle amplification (RCA) and single particle inductively coupled plasma mass spectrometry (spICP-MS) was proposed for the homogeneous detection of hepatitis B virus (HBV) deoxyribonucleic acid (DNA). In the presence of target DNA, long ssDNA possessing a large number of repeating sequence units was generated by RCA. DNA-labeled AuNP probes assembled into long chains based on complementary base pairing, further aggregating into large particles. Small Au NPs hardly produced pulse signals in spICP-MS; obvious pulse signals appeared in spICP-MS after the agglomeration of Au NPs caused by the addition of RCA products and spermidine. On the basis of this, the homogeneous detection of target DNA was realized by spICP-MS with high sensitivity. Under optimal conditions, the proposed method exhibited a good linear relationship between the frequency of the pulse signal of Au in spICP-MS and the concentration of target HBV DNA in the range of 10–2000 fmol L–1 (R = 0.997), the limit of detection was 5.1 fmol L–1, and the relative standard deviation was 3.7–6.8%. Recoveries of 94.2–108% were obtained for target DNA in spiked serum samples, demonstrating a good matrix tolerance ability for the method.
中文翻译：
基于单粒子电感耦合等离子体质谱的 HBV DNA 均相检测与滚环放大诱导金纳米粒子聚集
提出了一种基于滚环放大（RCA）和单粒子电感耦合等离子体质谱（spICP-MS）的高灵敏度、简便的乙型肝炎病毒（HBV）脱氧核糖核酸（DNA）均相检测方法。在靶DNA存在的情况下，RCA产生具有大量重复序列单元的长ssDNA。DNA 标记的 AuNP 探针基于互补碱基配对组装成长链，进一步聚集成大颗粒。小的 Au NPs 在 spICP-MS 中几乎不产生脉冲信号；RCA产物和亚精胺的加入导致Au NPs团聚后在spICP-MS中出现明显的脉冲信号。在此基础上，利用spICP-MS实现了目标DNA的均相检测，灵敏度高。在最优条件下，–1 ( R = 0.997)，检出限为5.1 fmol L –1，相对标准偏差为3.7-6.8%。加标血清样品中目标 DNA 的回收率为 94.2-108%，表明该方法具有良好的基质耐受能力。
更新日期：2022-07-05
中文翻译：
基于单粒子电感耦合等离子体质谱的 HBV DNA 均相检测与滚环放大诱导金纳米粒子聚集
提出了一种基于滚环放大（RCA）和单粒子电感耦合等离子体质谱（spICP-MS）的高灵敏度、简便的乙型肝炎病毒（HBV）脱氧核糖核酸（DNA）均相检测方法。在靶DNA存在的情况下，RCA产生具有大量重复序列单元的长ssDNA。DNA 标记的 AuNP 探针基于互补碱基配对组装成长链，进一步聚集成大颗粒。小的 Au NPs 在 spICP-MS 中几乎不产生脉冲信号；RCA产物和亚精胺的加入导致Au NPs团聚后在spICP-MS中出现明显的脉冲信号。在此基础上，利用spICP-MS实现了目标DNA的均相检测，灵敏度高。在最优条件下，–1 ( R = 0.997)，检出限为5.1 fmol L –1，相对标准偏差为3.7-6.8%。加标血清样品中目标 DNA 的回收率为 94.2-108%，表明该方法具有良好的基质耐受能力。
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