Aptasensor For Detection Research Articles

Ultralow Potential Cathodic Electrochemiluminescence Aptasensor for Detection of Kanamycin Using Copper Nanoribbons as Coreaction Accelerator.

An ultralow cathodic potential electrochemiluminescence (ECL) aptasensor was designed, employing DNA nanoribbon template self-assembly copper nanoclusters (DNR-CuNCs) as a novel coreaction accelerator within the luminol-H2O2 system for the sensitive detection of kanamycin (KANA). Mechanistic investigations revealed that the DNR-CuNCs preferred to generate highly active hydroxyl radicals by facilitating the reduction of the coreactant H2O2 under neutral pH conditions, consequently enhancing cathodic luminescence. By the strong π-π stacking effect of KANA aptamer and graphene as a signal modulation switch, DNR-CuNCs were displaced from the electrode surface due to the affinity of KANA and its aptamer, resulting in the inhibition of the luminol-H2O2 system and a decrease in the ECL signal. Under optimal experiments, the aptasensor demonstrated exceptional sensitivity in detecting KANA within the concentration range from 1 × 10-2 to 5 × 105 pg/mL, with the detection limit as low as 0.18 fg/mL. This innovative strategy provided a novel approach to designing effective ECL emitters for monitoring food safety.

Development of a rapid magnetic relaxation switching aptasensor for detection of saxitoxin using Fe3+/silver nanoparticles reaction system

Saxitoxin (STX) is a highly toxic marine bio-toxin, which has attracted great attention in the field of environmental and food safety. It is of crucial importance to develop efficient, sensitive and cost-effective methods for detecting STX. Here, we designed a magnetic relaxation switching (MRS) aptasensor for detection of STX based on FeCl3-regulated the aptamer-functionalized silver nanoparticles-coated magnetic nanoparticles (MNP@Ag NPs). In the light of the sensing effect of transverse relaxation time (T2) for optimizing, the etched time, the concentration of MNP@Ag NPs, the concentration of aptamers and incubation time were 15 min, 4 mg mL−1, 0.4 μM and 90 min, respectively. The MRS aptasensor showed good linearity amidst the sensing range of 0.2 μM to 1.0 μM and the limit of detection (LOD, S/N = 3) of 0.18 μM. Additionally, the MRS aptasensor demonstrated excellent sensitivity, reproducibility, and stability in clams and mantis shrimps. The recovery of STX ranged from 82.6 % to 101.5 % with low relative standard deviation (RSD) values, suggesting its effectiveness for detecting STX across a wide array of seafood samples.

An autocatalytic hybridization circuit-based FRET aptasensor for detection of low-abundant sulfameter in human serum

Exposure to antibiotics is considered a potential risk factor for human health. Yet, the extensive and cost-effective detection of low-abundant antibiotics in complex matrices remains a significant challenge. Herein, an aptamer and an autocatalytic hybridization circuit (AHC) were used to fabricate a fluorescence resonance energy transfer (FRET) platform to detect sulfameter (SME) in human serum. The AHC system comprised two mutually motivated hybridization chain reactions (HCR) modules, ultimately producing long-branched DNA copolymeric nanowires. This mutually reciprocal activation of two HCR modules enables continuous signal amplification, providing the AHC system with wide linear range and high sensitivity for the SME detection. Compared to the HCR-based aptasensor, the AHC-based aptasensor exhibited a wider linear range and improved sensitivity (3.3 times greater). Under optimal conditions, the fluorescent AHC-based aptasensor demonstrated a linear range (R2 was 0.996) from 0.5 to 2000 nM, with a low detection limit of 0.301 nM (S/N = 3). The fluorescent aptasensor was also validated by SME-spiked human serum samples, showing average recoveries ranging from 96.40 % to 109.30 %, with a relative standard deviation below 10.45 %. Furthermore, when tested on six human serum samples, the aptasensor results were consistent with those obtained from the commercial ELISA method. These findings demonstrate that the proposed aptasensor provides a promising approach for the practical monitoring of low-abundant SME in human serum.

Portable label-free electrochemical aptasensor for sensitive detection of paraquat herbicide mediated by oxygen reduction reaction

Paraquat (PQ) is a highly toxic herbicide that has been prohibited in almost 70 countries, but remains in use worldwide. Thus, routine on-site PQ monitoring is a key mechanism to ensure safety and efficiently enforce regulations. Herein, a label-free portable electrochemical aptasensor for the detection of PQ was developed by utilizing aptamer designed to specifically recognize PQ. The aptasensor employs square-wave voltammetry (SWV) to quantify PQ binding on the aptamer-functionalized electrode surface by tracking the downstream oxygen reduction reaction. It provided a detection range spanning from 0.01 to 100.0 μg mL−1 PQ with a limit of detection (LOD) of 8.9 ng mL−1. Validation against spiked tap water, pomegranate juice, and orange juice revealed recovery rate performances of 75 %–130 %. The aptasensor demonstrates promising feasibility for PQ detection in real-world applications, offering remarkable portability and operational simplicity. Notably, it can operate without supplementary redox agents, requiring only sample incubation and subsequent washing steps.

A novel red emission carbon quantum dots-based aptasensor for detection of potassium ions in vitro and in vivo and its application in plant imaging

The determination of potassium ion (K+) concentration in organisms is of particular importance. Herein, an efficient, sensitive, and eco-friendly fluorescence aptasensor for detecting potassium ions (K+) was designed based on the interaction of aptamer (K-Apt), complementary sequence of aptamer (C-Apt), red emission carbon quantum dots (R-CDs), and graphene oxide (GO). Red luminescent CDs modified with K+ aptamer(R-CDs@K-Apt) and C-Apt modified GO (C-Apt@GO) were used as the fluorescence donor and acceptor, respectively, in this aptasensor. K+ was detected sensitively using the fluorescence resonance energy transfer (FRET) method. The linear range of this aptasensor was 5.0 × 10−9–2.2 × 10−7 mol/L, and its detection limit was as low as 6.2 × 10−11 mol/L. Importantly, this aptasensor can be employed for quantitative K+ detection in serum because of the enhancement of its selectivity and biocompatibility by the modification of biomolecules including K-Apt and C-Apt. Furthermore, the aptasensor’s red emission at 625 nm enabled it to completely avoid the background fluorescence caused by chlorophyll in wheat seedlings, allowing it to display the K+ concentration of the wheat through plant imaging. These findings lay a foundation for the development of a novel detection technique for analyzing the distribution of K+ in the body fluids of animals and plants.

Generation of specific aptamers for constructing CoOOH nanozymes-based colorimetric aptasensor for detection of fleroxacin in milk

The misuse or abuse of fleroxacin (FLE) in livestock and aquaculture can result in elevated levels of FLE in animal-derived foods, posing significant health risks to humans. Detection of FLE has been relying on instrumental analysis methods that need expensive equipment and longer analysis time. Herein we report the first isolation of two specificity aptamers against FLE (FLE-9 and FLE-51) by capture-SELEX, characterized them well and explored the binding mechanism. Furthermore, FLE-9 was selected to construct a CoOOH@Apt based biosensor. The catalytic activity of the biosensor was evaluated using the Michaelis equation, revealing a Vmax of 4.87×10−8 M s−1 and a Km of 0.11 mM. These results suggest that the affinity of CoOOH@Apt is higher than that of CoOOH nanozymes. In the present of FLE, the aptamer FLE-9 specifically binds to FLE, distancing itself from CoOOH nanozymes and thereby reducing the catalytic activity of the biosensor. Finally, the label-free colorimetric aptasensor was successfully applied to detect FLE, with a detection limit of 4.26 ng/mL. The recovery rates ranged from 96.70 % to 101.70 %, with a relative standard deviation (RSD) below 3.02 %. This study presents promising probes and a novel strategy for the determination of FLE in milk samples, contributing to improved food safety monitoring.

Competitive electrochemical aptasensor for high sensitivity detection of liver cancer marker GP73 based on rGO-Fc-PANi nanocomposites

Golgi protein 73 (GP73) is a novel tumor marker in the early diagnosis and prognosis of hepatocellular carcinoma (HCC). Herein, a competitive electrochemical aptasensor for detecting GP73 was constructed using reduced graphene oxide-ferrocene-polyaniline nanocomposite (rGO-Fc-PANi) as the biosensing platform. The rGO-Fc-PANi had larger specific surface area, excellent conductivity and outstanding electroactive performance, which served as nanocarrier for GP73 aptamer (GP73Apt) binding and as redox nanoprobe for record electrical signal. Then, a complementary chain (cDNA) was fixed to the electrode by hybridization with GP73Apt. When GP73 was present, a competitive process happened among cDNA, GP73Apt and GP73, formed the GP73-GP73Apt stable chemical structure and made cDNA detach from the sensing electrode, resulting in enhancement of electrical signal. The difference in the corresponding peak current before and after the competition can be used to indicate the quantitative of GP73. Under optimal conditions, the DPV current response showed a good log-linear relationship with GP73 concentrations (0.001 ∼ 100.0 ng/mL) with a detection limit of 0.15 pg/mL (S/N = 3). It was successfully used for GP73 detection in human serum with RSDs ranging from 1.08 % to 6.96 %. Therefore, the aptasensor could provide an innovative technology platform and hold a great potential in clinical application.

Recent Developments on Optical Aptasensors for the Detection of Pro‐Inflammatory Cytokines with Advanced Nanostructures

AbstractIn the realm of immune response, pro‐inflammatory cytokines play a crucial role in regulating various physiological functions. Accurate measurement of these low‐molecular‐weight proteins is essential for understanding immune function, predicting diseases, and monitoring treatment effects. Optical aptasensors with advanced nanostructures, which utilize aptamers as bio‐probes, have emerged as a promising technology for cytokine detection, offering advantages over traditional antibody‐based nanobiosensors. Aptamers, single‐stranded nucleic acids with high specificity and affinity, enable cost‐effective mass production and consistent quality. Optical biosensors incorporating aptamers exhibit stability, resistance to environmental factors, and prolonged functionality. This review explores the current methodologies and advancements in optical aptasensors for cytokine detection, highlighting their potential as robust tools in diagnostics and therapeutics. Specifically, the applications of surface plasmon resonance and fluorescence techniques in aptasensors are discussed, focusing on the innovative approaches used to enhance sensitivity and specificity in cytokine detection. Notable examples of aptasensor designs utilizing nanoparticles, Förster resonance energy transfer, and amplification strategies are presented. These designs demonstrate high affinity, specificity, and improved sensitivity in detecting pro‐inflammatory cytokines such as interferon gamma. Overall, optical aptasensors show great promise in advancing the understanding of cytokine‐related disorders and enabling effective interventions.

Tag-free fluorometric aptasensor for detection of chromium(VI) in foods via SYBR Green I signal amplification and aptamer structure transition.

In response to growing concerns regarding heavy metal contamination in food, particularly chromium (Cr)(VI) contamination, this study presented a simple, sensitive and practical method for Cr(VI) detection. A magnetic separation-based capture-exponential enrichment ligand system evolution (SELEX) method was used to identify and characterize DNA aptamers with a high affinity for Cr(VI). An aptamer, Cr-15, with a dissociation constant (Kd) of 4.42 ± 0.44 μmol L-1 was obtained after only eight rounds of selection. Further innovative methods combining molecular docking, dynamic simulation and thermodynamic analysis revealed that CrO4 2- could bind to the 19th and 20th guanine bases of Cr-15 via hydrogen bonds. Crucially, a label-free fluorometric aptasensor based on SYBR Green I was successfully constructed to detect CrO4 2-, achieving a linear detection range of 60-300 nmol L-1 with a lower limit of detection of 44.31 nmol L-1. Additionally, this aptasensor was able to quantitatively detect CrO4 2- in grapes and broccoli within 40 min, with spike recovery rates ranging from 89.22% to 108.05%. The designed fluorometric aptasensor exhibited high selectivity and could detect CrO4 2- in real samples without sample processing or target pre-enrichment. The aptasensor demonstrated its potential as a reliable tool for monitoring Cr(VI) contamination in fruit and vegetable products. © 2024 Society of Chemical Industry.

Paper-based aptasensor for electrochemical detection of polyvalent antigen of dengue virus

ABSTRACT Dengue virus (DENV) is one of the most transmittable diseases. Numerous countries still face severe illness or deaths due to DENV. To date, there is no vaccine or effective drug available for this. Therefore, to overcome the burden of the disease, an early & sensitive diagnostic approach shows a main part in the management of dengue disease. The present research is aimed to construct PBAs i.e. paper-based aptasensor for the detection of all serotypes of DENV. The proposed biosensor has several advantages, including being simple, low-cost, reproducible, and disposable. The aptasensor uses a three-electrode system, with the working electrode containing a silver/zinc nanocomposite that was chemically synthesised. The characterisation of the zinc/silver nanocomposite were evaluated using UV-vis spectra, XRD, & FESEM. Aptamers were also employed to increase the sensor’s functioning. This aptamer functionalised sensor was employed to improvise the selectivity of the developed platform. The outcomes of aptasensor was evaluated via CV/LSV, which was verified using a potentiostat. The established paper-based aptasensor delivered ideal response and a broad linear variety of 0.1–1000 µg/ml for the DENV virus, having 0.1 µg/ml (LOD). The developed PBAs, or paper-based aptasensors, might be particularly useful in point-of-care diagnostic applications. This biosensor is capable of detecting polyvalent DENV-Antigen of 4 serotypes of DENV and hence, this research provides sensitive diagnosis of serotypes on a same platform and also offers viable diagnostic apparatus for rural places with limited resources, for people who can’t afford costly medical procedures and have a finite access to professional staff.

Green-synthesized chitosan‑carbon dot nanocomposite as turn-on aptasensor for detection and quantification of Leishmania infantum parasite

Leishmania is one of the most common diseases between human and animals, caused by Leishmania infantum parasite. Here, we have developed an ultra-selective turn-on fluorescent probe based on an aptamer and Chitosan-CD nanocomposite. The CD used in this study were synthesized using Quercus cap extract and a microwave-assisted approach. The Chitosan-CD nanocomposite was optimized using several microscopic and spectroscopic techniques to possess a bright fluorescence emission before adding aptamer and totally quenched fluorescence after addition of aptamer. The designed probe was proficient in the detection and quantification Leishmania infantum parasite by selective targeting of poly(A) binding protein (PABP) on the surface of the parasite. The designed fluorescent biosensor with high sensitivity, excellent selectivity, and a limit of detection (LOD) of 94 cells/mL of the Leishmania infantum parasite as well as a linear response in the ranges of 188–750 cells/mL and 3000–6000 cells/mL (R2 ≥ 0.98 for both linear ranges). Additionally, the selectivity of the designed probe was evaluated in the presence of different pathogenic species such as Trypanosoma brucei parasite and Staphylococcus aureus bacteria, as well as LiIF2α and LiP2a and BSA proteins as interference substances. The results of this study shows that using Chitosan-CD nanocomposite is a great strategy for developing selective turn-on probes with extraordinary accuracy and sensitivity in identifying Leishmania infantum parasite, especially in the early stages of the disease, and it is promising for the future clinical applications.

A photothermal aptasensor based on rolling circle amplification-enriched DNAzyme for portable detection of ochratoxin A in grape juice

Aptasensors for detection of ochratoxin A (OTA) have been extensively studied, but the majority of them require costly and large-scale equipment as signal readers. Herein, a photothermal aptasensor capable of portable detection of OTA through a thermometer was developed on basis of aptamer structural switching and rolling circle amplification (RCA)-enriched DNAzyme. Oligonucleotides and alkaline phosphatase (ALP) modified magnetic beads were prepared. The binding of aptamers to OTA led to the release of ALP labeled complementary DNA. After magnetic separation, ALP catalyzed the padlock dephosphorylation, inhibiting the subsequent RCA reaction. This process converted the OTA concentration into the amount of the photothermal reagent oxTMB produced from the catalytic reaction induced by RCA-enriched DNAzyme. Under the optimal conditions, the detection limit (LOD) of this aptasensor was 2.28 nM in a clean buffer, while the LOD reached 2.43 nM in 2 % grape juice. The good performance of the photothermal aptasensor makes it possible to measure OTA pollution in low resource environments.

An electrochemical aptasensor for methylamphetamine rapid detection by single-on mode based on competition with complementary DNA

A simple and rapid electrochemical sensing method with high sensitivity and specificity of aptamers was developed for the detection of methylamphetamine (MAMP). A short anti-MAMP thiolated aptamer (Apt) with a methylene blue (MB) probe at 3ʹ-end was immobilized on the surface of a gold electrode (MB-Apt-S/GE). The electrochemical signal appeared when MAMP presenting in the sample solution competed with cDNA for binding with MB-Apt-S. Under optimized conditions, the liner range of this signal-on electrochemical aptasensor for the detection of MAMP achieved from 1.0 to 10.0 nmol/L and 10.0–400 nmol/L. LOD 0.88 nmol/L were obtained. Satisfactory spiked recoveries of saliva and urine were also obtained. In this method, only 5 min were needed to incubate before the square wave voltammetry (SWV) analysis, which was much more rapid than other electrochemical sensors, leading to a bright and broad prospect for the detection of MAMP in biological sample. This method can be used for on-site rapid detection on special occasions, such as drug driving scenes, entertainment venues suspected of drug use, etc.

Label-free electrochemical aptasensor for detection of Acinetobacter baumannii: Unveiling the kinetic behavior of reduced graphene oxide v/s graphene oxide

In this study, we introduce an electrochemical aptasensor for the sensitive detection of Acinetobacter baumannii. This method, based on impedance and current analysis, represents a first-of-its kind platform in the field of A. baumannii detection. We employed a screen-printed carbon electrode modified with both graphene oxide (GO) and reduced graphene oxide (rGO) as the immobilization platform, and the bacteria specific aptamer was employed as the biorecognition element. The aptamer selectively interacts with A. baumannii, causing a decrease in the peak current of the redox complex because the binding biomolecules on the electrode surface hinder the electron transfer of the redox probe. This initial point-of-care-compatible rGO electrochemical aptasensor demonstrated an exceptional sensing performance with an impressive limit of detection of 10 CFU/mL for detecting A. baumannii. This enhanced sensitivity can be attributed to several key factors including structural defects during reduction, increased surface area and improved electron transfer kinetics. The findings of this study highlight the potential of the developed aptasensor as a rapid and reliable detection platform for various pathogenic diseases. By further ensuring seamless integration into clinical practice, these label-free graphene-based aptasensors, with their π-π interactions and other favorable attributes, hold great promise for sensitive and specific pathogen detection, thereby contributing to advancements in healthcare diagnostics.

Portable smartphone-based aptasensor for nitrofuran detection

A portable electrochemical aptasensor for the detection of nitrofuran (NF) was developed. Aptamer-conjugated gold nanoparticles (Apt-AuNPs) were immobilized on a chitosan (CS) thin film electrodeposited on a screen-printed carbon electrode (SPCE). The modified electrode was connected to a smartphone-controlled near-field communication (NFC) potentiostat. The electrochemical aptasensor determined NF concentration using cyclic voltammetry (CV) and chronoamperometry (CA) techniques. Under optimal conditions, the developed aptasensor exhibited high sensitivity (11.26 ± 0.38 μA L/μg), a wide linear range (0.005 to 0.08 μg/L and 0.08 to 4.0 μg/L), and a low detection limit (0.004 μg/L). Our construction demonstrated excellent stability for NF, with a reduction in current of less than 10 % even after 330 days. Reproducibility (%RSD = 2.19 %) was good, and the device was portable, simple, and selective. Therefore, this aptasensor platform was applied as a smartphone-based wireless NFC potentiostat for NF detection in shrimp pond water and shrimp samples.

CRISPR-Cas12a-based colorimetric aptasensor for aflatoxin M1 detection based on oxidase-mimicking activity of flower-like MnO2 nanozymes

Given the highly mutagenic and carcinogenic nature of Aflatoxin M1 (AFM1), the quantity assessment of AFM1 residues in milk and dairy products is necessary to maintain consumer health and food safety. Herein, CRISPR-Cas12a-based colorimetric aptasensor was developed using the catalytic activity of flower-like nanozymes of MnO2 and trans-cleavage property of CRISPR-Cas12a system to quantitatively detect AFM1. The basis of the developed colorimetric aptasensor relies on whether or not the CRISPR-Cas12a system is activated, as well as the contrast in oxidase-mimicking capability exhibited by flower-like MnO2 nanozymes when AFM1 is absent or present. When AFM1 is not present in the sample, single-stranded DNA (ssDNA) is degraded by the activated CRISPR-Cas12a, and the solution turns into yellow due to the catalytic activity of the nanozymes. While, in the attendance of AFM1, ssDNA degradation does not occur due to the inactivation of the CRISPR-Cas12a. Therefore, with the adsorption of the ssDNA on the MnO2 nanozymes, their catalytic activity decreases, and the solution color becomes pale yellow due to less oxidation of the chromogenic substrate. In this aptasensor, the relative absorbance changes increased linearly from 6 to 160 ng L−1, and the detection limit was 2.1 ng L−1. The developed aptasensor displays a selective detection performance and a practical application for quantitative analysis of AFM1 in milk samples. The results of the introduced aptasensor open up the way to design other selective and sensitive aptasensors for the detection of other mycotoxins by substitution of the used sequences.

Probing aptamer-mucin 1 binding events on polydopamine@carbon nanotubes modified cellulose paper interface using speckle pattern analysis for label free aptasensing

Herein, we have developed a cellulose paper-based aptasensing protocol by employing dynamic laser speckle (DLS) analysis for direct and label-free probing of the binding interface. Cellulose filter paper surface was modified by using the mussel adhesion chemistry of polydopamine modified carbon nanotubes (PDA@CNTS). The mussel chemistry resulted in strong adhesion and binding of PDA@CNTs on cellulose paper surface. The modified surface was subsequently employed in the construction of label free aptasensor for detection of cancer biomarker mucin 1 (MUC1). Taking advantage of DLS technique, it provided us with sensitive and accurate analysis of target biomarker MUC1 by the intrinsic randomness of patterns generated from cellulose based paper surface. The developed aptasensor was evaluated using DLS at various fabrication steps, where the scattered laser beams from these samples were collected and analyzed based on the obtained speckle patterns. Under the optimal experimental conditions, paper-based aptasensor achieved sensitive and selective detection of MUC1, exhibiting excellent linear range (15–100) µM with a limit of detection of 1 µM. This work presents a novel study of promising laser speckle paper-based aptasensor, providing new insights for the development of low-cost, sensitive, and portable paper-based analytical tools for biological applications.

Multiplexed aptasensor for detection of acute myocardial infraction (AMI) biomarkers

Schematic illustration of the developed aptasensor for multiplex detection of AMI biomarkers.

Sensitive SERS aptasensor for histamine detection based on Au/Ag nanorods and IRMOF-3@Au based flexible PDMS membrane

BackgroundHistamine is a kind of biogenic amine with strong toxicity and potential carcinogenicity. Many traditional methods of detecting histamine have the disadvantages of cumbersome detection steps, expensive equipment, and high professional requirements for staff. In contrast, SERS has become the preferred method for quantitative analysis of histamine because of rich fingerprint information, rapidity and economy. However, most of SERS substrates still have technical problems, such as poor stability, low sample collection rate, and detection efficiency. Therefore, there is a great need for new strategies to develop high-performance SERS substrates based sensors. ResultsIn our study, a sensitive SERS aptasensor for the detection of histamine was synthesized. The assembly was formed between IRMOF-3@Au/PDMS (flexible SERS substrate) and AuNR-DTNB@Ag-HA apt (Raman signal probe with both the target capture ability) via π-π stacking interaction from HA aptamer and IRMOF-3. Consequently, the SERS signal of the assembly derived from DTNB reached highest due to the synergistic enhancement effect by AuNR@Ag and IRMOF-3@Au. Meanwhile, HA aptamer can specifically capture histamine, therefore histamine addition competitively bound to the probe, leading to a corresponding decrease in the DTNB signal value on the SERS substrate. The SERS intensity at 1331 cm−1 presented a good linear relationship towards the logarithmic value of histamine concentrations ranging from 0.0001 mg/L to 400 mg/L (R2 = 0.990) with the LOD of 3.6 × 10−5 mg/L. Furthermore, the application in wine samples demonstrated the accuracy and applicability of the developed sensor. SignificanceThis method effectively improves substrate stability, detection sensitivity and signal response immediacy to amplify the SERS sensor, thus satisfying the histamine detection requirements of various systems. According to this aptasensor design, our strategy can be extended to create other MOF-based SERS substrates for accurately detecting relative targets to ensure food safety.

Devising a colorimetric aptasensor for detection of basal stem rot-associated RNA marker during early Ganoderma boninense infection in oil palm

Basal stem rot (BSR) is a detrimental disease in oil palm (Elaeis guineensis) due to the host's lack of symptoms during the early phase of infection. Early detection of BSR at non-symptomatic phase is feasible via molecular-based approaches and has become the major interest among researchers. Our previous study had identified a differentially expressed gene encoding auxin efflux carrier component 5-like (EgPIN5) at 11 day-post-inculation of Ganoderma boninense (symptomless early stage of infection) on oil palm seedlings. We further evaluated the gene expression on samples collected from an oil palm plantation at different BSR stages (symptomless, mild and severe). Our findings revealed that EgPIN5 was expressed in symptomless group but was not expressed in either mild or severe BSR groups. Next, we developed a sandwich-type aptamer-based lateral flow assay (LFA) against EgPIN5 RNA by adapting the versatile DNA-functionalized gold nanoparticles for colorimetric detection. The LFA strip demonstrated visible colorimetric signal of EgPIN5 RNA at 12.5 ng/μL with limit of detection at 2.45 ng/μL using a scanning device. Based on our consistent results, we presented EgPIN5 as a diagnostic biomarker for symptomless BSR infection in oil palm. Utilization of LFA assay strip for practical EgPIN5 RNA detection is hoped to encourage establishment of a fast, cheap, and sensitive point-of-care detection system in plant biology.