The Dual-Response–Single-Amplification Fluorescent Nanomachine for Tumor Imaging and Gastric Cancer Diagnosis

Abstract

Gastric cancer (GC) is one of the most common tumors worldwide and is the leading cause of tumor-related mortality. Traditional biomarkers and screening methods cannot meet the clinical demands. There is an urgent need for highly sensitive diagnostic markers as well as accurate quantification methods for early gastric cancer (EGC) screening. Here a dual-target cooperatively responsive fluorescent nanomachine by the innovative application of two targets� responsive strand migration system with a single-amplificationcycle element was developed for the simultaneous detection of GC biomarkers miR-5585-5p and PLS3 mRNA, which were selected by next-generation sequencing and RT-qPCR. It was also an RNA extraction-free, PCR-free, and nonenzymatic biosensor to achieve tumor cell imaging and serum diagnosis. Requiring only a 20 μL serum sample and 20 min incubation time, the nanomachine achieved an ultrasensitive detection limit of fM level with a broad linear range from fM to nM. More importantly, a higher AUC value (0.884) compared to the clinically used biomarker CA 72-4 was obtained by the nanomachine to distinguish GC patients successfully. Notably, for the key concerns of diagnosis of EGC patients, the nanomachine also achieved a satisfactory AUC value of 0.859. Taken together, this work has screened and obtained multiple biomarkers and developed a fluorescent nanomachine for combination diagnosis of GC, providing an ingenious design of a functionalized DNA nanomachine and a feasible strategy for the transformation of serum biomarkers into clinical diagnosis.

Can Nano Yield Big Insights? Oligonucleotide-Based Biosensors in Early Diagnosis of Gastric Cancer

“…Multiplexing is pivotal for biosensors as it enables the simultaneous detection of multiple analytes, enhancing efficiency and providing comprehensive diagnostic information [89]. Zhang et al (2023) introduced an innovative dual-target responsive fluorescent nanomachine for the simultaneous detection of miR-5585-5p and PLS3 mRNA [83]. Guided by advanced techniques such as next-generation sequencing, the nanomachine operated without the need for RNA extraction or PCR, ensuring simplicity.…”

A Universal Strategy for Enhancing the Circulating miRNAs’ Detection Performance of Rolling Circle Amplification by Using a Dual-Terminal Stem-Loop Padlock

“…Compared to traditional protein biomarkers, nucleic acid biomarkers can respond to the disease state at the transcriptional level, which is the most promising field for early precision diagnosis and treatment. , Rolling circle amplification (RCA) is one of the most promising isothermal amplification techniques and has been used in a wide range of fields, such as nanomaterials, biochemistry, and molecular biology. − In nucleic acid detection, padlock probes are commonly used to specifically identify nucleic acid targets and be connected into circular templates, which is then followed by RCA for signal amplification and output, which is commonly involved in molecular diagnosis of various diseases. , Conventional padlock probes are linear DNA sequences that typically contain two target recognition regions at both terminals and a functional region in the middle. By changing the target recognition regions, padlock probes can target different nucleic acid molecules .…”