Microarray can provide genomic and proteomic information for several biological targets at the same time, which is a significant progress because they provide more complete and reliable information and simplify the whole analysis scheme. Creative Biolabs provides this microsphere-based multiplex immunoassay for simultaneous detection of protein and DNA in a single test.
Background of Microsphere-based Multiplex Immunoassay
Microsphere assays have become an important form of multiple analysis such as genotyping, gene expression and protein immunoassay. Different microsensors can be analyzed simultaneously by chemical, spectrometric or physical means to encode each type of probe functionalized microspheres. A rolling circle amplification (RCA) microarray based on high-density microspheres has been developed for simultaneous detection of protein and DNA in a single test. With this new method of detecting proteins and protein related target DNA, medical conditions can be diagnosed by a rapid analysis, rather than using different tests to detect different disease markers.
Introduction of RCA
RCA is an alternative amplification method for improving the sensitivity of DNA quantification, DNA mutation detection and array-based sandwich immunoassay. Due to its strict requirements for the matching of the stringent strands, RCA is a simple, highly sensitive and specific amplification method that can produce high-efficiency amplification.
RCA products contain thousands of repeat sequences that complement fluorescent labeled signal probes. Because RCA can be used as a signal generation method for antibody detection, it can also be compatible with the sandwich immunoassay format. The integrity of the antibody-antigen complex was maintained during isothermal amplification, and the amplified DNA products could be covalently or connected to the detection antibody through the avidin biotin bridge.
Fig.1 Illustration of RCA. Distributed under CC BY-SA 4.0, from Wiki, without modification.
Services in Creative Biolabs
Our microsphere-based multiplex immunoassay platform can realize the combined detection of nucleic acid sequences and proteins in one detection. This assay only generates a signal when both analytes are present simultaneously. Due to the high amplification efficiency of RCA and the high binding capacity of the microsphere surface, the detection limit of protein and DNA can reach fM and pM levels, respectively.
By collecting a wide range of catalog and non-catalog nucleic acids, antibodies and proteins, our team can quickly assess the feasibility of the target and add it to the existing microsphere array, ultimately reducing your time to obtain data and minimizing the sample size required. Our customized services include array optimization, microsphere modification and customized analysis and development.
If you are interested in our service, please contact us for your exclusive solution.
Published Data
1. Multiplex Microsphere Immunoassay for Detecting Antibodies to Highly Pathogenic Viruses
Fig.2 Validation of MMIA with serum samples from virus-infected non-human primates and human individuals with known infection status.1
This study developed a multiplex microsphere immunoassay (MMIA) for detecting IgG antibodies targeting the nucleoproteins (NPs) of CCHFV, DOBV, RVFV, EBOV, and MARV. Upon optimization, the assay demonstrated specific binding to each NP, tested on sera from humans and non-human primates with confirmed infection status. The assay's potential for serosurveillance was validated by analyzing 129 human serum samples from Guinea and comparing them with 88 samples from the German blood bank. The MMIA showed good agreement with commercial ELISA and IIFT methods, with significantly higher binding to EBOV and MARV NPs in the Guinea samples. Additionally, the assay was successfully adapted to detect antibodies in bats inoculated with EBOV and MARV virus-like particles, showcasing its versatility for monitoring both wildlife and human populations. This high-throughput, broadly reactive assay can be used to screen for antibodies against multiple highly pathogenic viruses.
2. Microsphere Multiplex Assays and Mixture Models for Tropical Disease Serosurveillance
Fig.3 Evaluation of the microsphere-based multiplex assay.3
This study developed a microsphere-based multiplex serological assay to measure IgG antibody levels for six infectious diseases: Leishmania donovani, Toxoplasma gondii, Entamoeba histolytica, HIV, Wuchereria bancrofti, and Vibrio cholerae. Researchers conducted a serological survey in two health and demographic surveillance areas in coastal and western Kenya, randomly selecting 4,600 individuals based on sex and age, with 3,411 agreeing to participate. Mathematical analyses of participants' responses to each antigen, combined with the serostatus of controls, suggested that this assay could be useful for monitoring infections, particularly HIV, toxoplasmosis, filariasis, leishmaniasis, and amebiasis. The system allows for the addition of pathogens and antigens of interest, offering a flexible approach for future monitoring programs tailored to specific needs in Africa.
References
- Surtees, Rebecca, et al. "Development of a multiplex microsphere immunoassay for the detection of antibodies against highly pathogenic viruses in human and animal serum samples." PLoS Neglected Tropical Diseases 14.10 (2020): e0008699. Distributed under Open Access license CC0 1.0, without modification.
- Fujii, Yoshito, et al. "Serological surveillance development for tropical infectious diseases using simultaneous microsphere-based multiplex assays and finite mixture models." PLoS neglected tropical diseases 8.7 (2014): e3040. Distributed under Open Access license CC BY 4.0, without modification.
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