A comprehensive review of aptamer screening and application for lateral flow strip: Current status and future perspectives.

The detection of biomarkers is of great significance for medical diagnosis, food safety, environmental monitoring, and agriculture. However, bio-detection technology at present often necessitates complex instruments, expensive reagents, specialized expertise, and prolonged procedures, making it challenging to fulfill the demand for rapid, sensitive, user-friendly, and economical testing. In contrast, lateral flow strip (LFS) technology offers simple, fast, and visually accessible detection modality, allowing real-time analysis of clinical specimens, thus finding widespread utility across various domains. Within the realm of LFS, the application of aptamers as molecular recognition probes presents distinct advantages over antibodies, including cost-effectiveness, smaller size, ease of synthesis, and chemical stability. In recent years, aptamer-based LFS has found extensive application in qualitative, semi-quantitative, and quantitative detection across food safety, environmental surveillance, clinical diagnostics, and other domains. This review provided a concise overview of different aptamer screening methodologies, selection strategies, underlying principles, and procedural, elucidating their respective advantages, limitations, and applications. Additionally, we summarized recent strategies and mechanisms for aptamer-based LFS, such as the sandwich and competitive methods. Furthermore, we classified LFSs constructed based on aptamers, considering the rapid advancements in this area, and discussed their applications in biological and chemical detection. Finally, we delved into the current challenges and future directions in the development of aptamer and aptamer-based LFS. Although this review was not thoroughly, it would serve as a valuable reference for understanding the research progress of aptamer-based LFS and aid in the development of new types of aptasensors.

Gold-based paper for antigen detection of monkeypox virus.

In 2022, the outbreak of the monkeypox virus occurred in many non-endemic countries, and the World Health Organization (WHO) assessed that this outbreak was "atypical". The establishment of a rapid and effective assay that can be used for the early diagnosis of monkeypox virus infection is crucial for outbreak prevention and control. In this study, the monkeypox virus A29 protein and the homologous vaccinia virus A27 protein and cowpox virus 162 protein were expressed in Escherichia coli BL21 for screening. We synthesized the monkeypox virus A2917-49 peptide as the immunogen and obtained 25 monoclonal antibodies (mAbs) against the A29 protein using mouse hybridoma techniques. Then an immunochromatographic test strip method for detecting A29 was established. The strips utilizing mAb-7C5 and 5D8 showed the best sensitivity and lowest limit of detection: 50 pg mL-1 for purified A29 and specificity tests showed that the strips did not cross-react with other orthopox viruses (vaccinia virus or cowpox virus) as well as common respiratory pathogens (SARS-CoV-2, influenza A and influenza B). Therefore, this method can be used for early and rapid diagnosis of monkeypox virus infection by antigen detection.

Fluorescent microsphere-based lateral-flow immunoassay for rapid and sensitive determination of eugenols.

In this study, a sensitive monoclonal antibody (mAb) 1B5 against eugenols was prepared based on a novel hapten. Based on this mAb, a paper-based lateral-flow immunoassay (LFIA) was developed using Eu-fluorescent microspheres sensor, that could achieve qualitative and quantitative detection of eugenols within 10 min. Results showed colorimetric values observed by the naked eye were 12.3 µg/kg, 12.3 µg/kg, 37 µg/kg and 111 µg/kg for eugenol, isoeugenol, methyl eugenol, and methyl isoeugenol, respectively, in both water and fish samples. For quantitative detection of eugenol, isoeugenol, methyl eugenol and methyl isoeugenol, the detection ranges were 4.49-48.4 µg/kg, 6.02-66.8 µg/kg, 16.5-150 µg/kg and 47.9-710 µg/kg in water, and 3.9-30.9 µg/kg, 5.9-62.6 µg/kg, 16.7-255 µg/kg, and 44.5-890 µg/kg in fish, respectively. The recovery test and detection in fish demonstrated the reliability of the LFIA in real samples. Therefore, the developed LFIA produced a promising alternative tool for the rapid on-site detection of eugenols.

Fluorescent strip sensor for rapid and ultrasensitive determination of fluoroquinolones in fish and milk.

The synthetic antibiotics fluoroquinolones are popular due to their good antibacterial performance and low price, but the risk to human health caused by their residues has attracted great attention. In this study, an ultra-sensitive mAb, 4D7, was prepared with an IC50 of 0.027 ng mL-1 to norfloxacin (NOR) and cross-reactivity of 19.7-47.7% to lomefloxacin (LOM), pefloxacin (PEF), ofloxacin (OFL), enrofloxacin (ENR), ciprofloxacin (CIP), and danofloxacin (DAN). Based on mAb 4D7 and Eu-fluorescent microspheres, a rapid and sensitive immunochromatographic strip was developed for the detection of fluoroquinolone residues in fish and milk. The detection ranges (IC20-IC80) of the strip for the detection of NOR, PEF, LOM, OFL, ENR, CIP and DAN were 0.19-1.1 µg kg-1, 0.39-2.1 µg kg-1, 0.5-2.6 µg kg-1, 0.43-3.3 µg kg-1, 0.61-3.5 µg kg-1, 0.69-5.5 µg kg-1, 0.52-3.4 µg kg-1 in fish, and 0.027-0.19 µg kg-1, 0.049-0.34 µg kg-1, 0.069-0.39 µg kg-1, 0.06-0.41 µg kg-1, 0.089-0.65 µg kg-1, 0.12-0.81 µg kg-1, 0.091-0.52 µg kg-1 in milk, respectively. The recovery rates in spiked sample tests were 88.6-113.6% with a coefficient of variation less than 8.4%. Thus the newly-developed strip was sensitive and reliable for rapid on-site detection of fluoroquinolone residues in fish and milk.

Quantitative and rapid detection of spinosad and spinetoram by a gold nanoparticle-based immunostrip.

Spinosad (SPI) and spinetoram (Et-SPI) are currently among the most popular new insecticides because of their high efficiency and low toxicity. However, excessive residues in food still pose a potential risk to public health. Therefore, it is necessary to strengthen residue monitoring of the two insecticides based on a simple and rapid method. In this study, a highly sensitive mAb (6G9) against SPI and Et-SPI was prepared using the hapten SPI-HS and used to develop a colloidal gold nanoparticle-based immunochromatographic strip for the detection of SPI and Et-SPI in samples. The quantitative ranges of the developed strip for SPI and Et-SPI were 8.93-1633 ng g-1 and 20.3-3555 ng g-1 in rice, 32.6-785 ng g-1 and 79.3-1862 ng g-1 in tea, and 9.66-360 ng g-1 and 23.9-931 ng g-1 in onions, respectively. In addition, recovery rates ranged from 85.7% to 112.7% with a coefficient of variation <9.5%. Therefore, our developed method was sensitive and valid as a quantitative tool for the rapid monitoring of SPI and Et-SPI in foods.

Immunochromatographic assays for ultrasensitive and high specific determination of enrofloxacin in milk, eggs, honey, and chicken meat.

Enrofloxacin, a veterinary antibiotic that persists in food, poses a risk to human health. Here, a monoclonal antibody against enrofloxacin, 1H12, was prepared based on the hapten ENR-1, and showed excellent sensitivity with a 50% inhibitory concentration (IC50) of 0.03 ng/mL. Using this antibody, 2 lateral-flow immunochromatographic assays were developed for determination of enrofloxacin in egg, milk, honey, and chicken meat samples. The detection ranges (IC20-IC80) were 0.16-0.82 ng/g, 0.24-1.8 ng/g, 0.25-3.6 ng/g, and 0.61-3.9 ng/g by colloidal gold-immunochromatographic sensor (CG-ICS) analysis, and 0.022-0.42 ng/g, 0.054-0.42 ng/g, 0.069-1.4 ng/g, and 0.19-2.2 ng/g by Eu-fluorescence-immunochromatographic sensor (EF-ICS) analysis. The intraassay and interassay recovery rates were 88.9 to 108.5% with coefficients of variation of 1.3 to 7.0% by CG-ICS analysis, and 88.6 to 113.6% with coefficients of variation of 1.3 to 8.1% by EF-ICS analysis. Thus, our newly developed ICS are sensitive and reliable, providing an option for rapid quantitative detection of enrofloxacin in food samples.

Rapid quantitative determination of fentanyl in human urine and serum using a gold-based immunochromatographic strip sensor.

Fentanyl is a typical opioid that is used in surgical anesthesia. However, when abused, fentanyl can lead to addiction and even death. To better control the use of fentanyl, it is necessary to develop rapid and sensitive detection methods. In this study, an ultrasensitive monoclonal antibody (mAb) was prepared and used to develop an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) and a colloidal gold-based immunochromatographic strip (CG-ICS) for the analysis of fentanyl in urine and serum. Under optimum conditions, the anti-fentanyl mAb belonging to the subtype of IgG2b showed a half-maximal inhibitory concentration (IC50) of 0.11 ng mL-1 and a linear range of detection of 0.020-0.50 ng mL-1. Fenanyl-spiked original urine and serum diluted eight times were used for the analysis of fentanyl by ic-ELISA and CG-ICS. IC50 from the standard curves was 0.46 ng mL-1 for urine and 2.6 ng mL-1 for serum in ic-ELISA and 1.6 ng mL-1 for urine and 6.27 ng mL-1 for serum in CG-ICS. The recovery test revealed that the ic-ELISA and CG-ICS, with a recovery rate of 87.0-108.4% and a coefficient of variation of 3.3-10.9%, were the same reliable tools as the liquid chromatography tandem mass spectrometry for fentanyl analysis in real samples.

Chemical composition and evaluation of antioxidant activities, antimicrobial, and anti-melanogenesis effect of the essential oils extracted from Dalbergia pinnata (Lour.) Prain.

ETHNOPHARMACOLOGICAL RELEVANCE: Dalbergia pinnata (Lour.) Prain (D. pinnata) is a plant widely distributed in tropical and subtropical regions of Asia, Africa, and the Americas. In humans, it is used in the prevention and treatment of diseases such as respiratory system, digestive system, cardiovascular and cerebrovascular diseases. AIM OF THE STUDY: This study was aim to evaluate chemical composition, antioxidant activities, antimicrobial, and anti-melanogenesis properties of Essential oils (EO) from D. pinnata. MATERIALS AND METHODS: In this paper, the EO of D. pinnata were extracted using the supercritical CO2 extraction method and purified by molecular distillation. The volatile compounds of EO were characterized using Gas Chromatography-Mass Spectrometer (GC-MS). The antioxidant activities were evaluated by the methods of 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging, 2, 2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging assays. And two Gram-positive bacteria, three Gram-negative bacteria and a fungus were employed to evaluate the antimicrobial activity. The zebrafish was used as experimental model to evaluate the anti-melanogenesis effect of the EO from D. pinnata. RESULTS: The EO of D. pinnata were obtained in a yield of 4.75% (v/w) calculated on dry weight basis. 14 volatile compounds could be detected and the predominant components include elemicin (91.06%), methyl eugenol (3.69%), 4-allyl-2,6-dimethoxyphenol (1.16%), and whiskey lactone (0.55%). The antioxidant assay showed that the EO could scavenge DPPH (IC50 values of 0.038 mg/mL) and ABTS (IC50 value of 0.032 mg/mL) free radical, indicating that the EO had strong antioxidant activity. The results of antimicrobial test showed that Staphylococcus aureus was most sensitive to EO with minimal inhibitory concentration (MIC) of 0.78 µL/mL, followed by Streptococcus pyogenes (6.25 µL/mL) and Candida albicans (12.5 µL/mL). Gram-negative strains, including Escherichia coli, Pseudomonas aeruginosa and Salmonella typhimurium, were slightly affected by the EO. Additionally, EO from D. pinnata could reduce tyrosinase activity and melanin synthesis of zebrafish embryos in dose-dependent manner. And EO exhibited the more obvious anti-melanogenic effect compared with the positive control arbutin at the same dose (30 mg/L). CONCLUSIONS: Our results validated the main activities attributed to D. pinnata for its antimicrobial and antioxidant. In addition, the potent inhibitory impacts of EO on the pigmentation provides a theoretical basis for the in-depth study of the EO from D. pinnata and their application in pharmaceutical and cosmetic industries.

Development of Indirect Competitive Enzyme-Linked Immunosorbent and Immunochromatographic Strip Assays for Tiamulin Detection in Chicken.

Tiamulin (TML) is a diterpenoid antibiotic used in animals. In this study, a gold nanoparticle immunochromatographic strip assay and an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) were developed to detect the residue of TML in chicken. TML aminobutyric was synthesized and conjugated to keyhole limpet hemocyanin by mixed anhydride method as immunogen, whereas TML was connected to ovalbumin with 1,1'-carbonyldiimidazole as coating antigen. Under optimized conditions, the ultrasensitive monoclonal antibody-based ic-ELISA exhibited a half-maximal inhibitory concentration (IC50) value of 0.36 ng/mL with a working range of 0.14-0.9 ng/mL for TML. A rapid and sensitive immunochromatographic strip assay was developed with a TML cutoff value of 2.5 ng/mL. On the basis of these results, both developed methods are useful for TML detection in chicken.