Highly Sensitive and Specific Mass Spectrometric Platform for miRNA Detection Based on the Multiple-Metal-Nanoparticle Tagging Strategy

Abstract

The multiple-metal-nanoparticle tagging strategy has generally been applied to the multiplexed detection of multiple analytes of interest such as microRNAs (miRNAs). Herein, it was used for the first time to improve both the specificity and sensitivity of a novel mass spectroscopic platform for miRNA detection. The mass spectroscopic platform was developed through the integration of the ligation reaction, hybridization chain reaction amplification, multiple-metal-nanoparticle tagging, and inductively coupled plasma mass spectrometry. The high specificity resulted from the adoption of the ligation reaction is further enhanced by the multiple-metal-nanoparticle tagging strategy. The combination of hybridization chain reaction amplification and metal nanoparticle tagging endows the proposed platform with the feature of high sensitivity. The proposed mass spectrometric platform achieved quite satisfactory quantitative results for Let-7a in real-world cell line samples with accuracy comparable to that of the real-time quantitative reverse-transcriptase polymerase chain reaction method. Its limit of detection and limit of quantification for Let-7a were experimentally determined to be about 0.5 and 10 fM, respectively. Furthermore, due to the unique way of utilizing the multiple-metal-nanoparticle tagging strategy, the proposed platform can unambiguously discriminate between the target miRNA and nontarget ones with single-nucleotide polymorphisms based on their response patterns defined by the relative mass spectral intensities among the multiple tagged metal elements and can also provide location information of the mismatched bases. Its unique advantages over conventional miRNA detection methods make the proposed platform a promising and alternative tool in the fields of clinical diagnosis and biomedical research.

A Novel Ratiometric Electrochemical Biosensor Using Only One Signal Tag for Highly Reliable and Ultrasensitive Detection of miRNA-21

“…MicroRNAs (miRNAs), as representative nucleic acid biomarkers, are a group of endogenous, noncoding, single-stranded RNAs that perform an incomparable role in regulating genetic expression and cellular processes involving proliferation, differentiation, and apoptosis. − To date, many research studies have proven that various major diseases are associated with the abnormal expression of miRNA. − Therefore, considerable efforts have been devoted to establish sensitive and specific miRNA sensing strategies to develop the early diagnosis of cancers and targeted anticancer drugs. − Nevertheless, owing to the low abundance, small size, and high sequence homology of miRNA fragments, most biosensors suffered from different troubles in practical applications, involving poor robustness, reproducibility, accuracy, and reliability. − …”

Atomic spectrometry update: review of advances in the analysis of clinical and biological materials, foods and beverages

“…[186][187][188][189] Sandwich hybridization on streptavidin coated microtitre plates was used to detect three target miRNAs with three NP probes (Ag, Au and Pt), quantied by ICP-MS. 186 Improved selectivity, sensitivity and dynamic range was achieved in a multiple-metal-NP-tagging method with hybridization chain reaction amplication. 187 Here, three metal NP probes (Ag, Au and Pd) interacted with one target miRNA and ICP-MS response patterns were interrogated to enhance selectivity compared with other miRNA detection methods. Kang et al 188 presented a multiplexed assay using three MNAzymes (multicomponent nucleic acid enzyme) paired with three target miRNAs and three lanthanide tags with MB-DNA probes.…”

Real-Time Biosensor Platform Based on Novel Sandwich Graphdiyne for Ultrasensitive Detection of Tumor Marker

“…MicroRNAs (miRNAs) are emerging as an essential player in tissue differentiation, proliferation, and hematopoiesis. − However, their clinical detection remains a great challenge due to low content and highly similar genetic sequences. At present, the reported methods still have inherent defects, , and thus, new detection methods are urgently needed.…”