Development and evaluation of a Luminex multiplex serology assay to detect antibodies to bovine herpes virus 1, parainfluenza 3 virus, bovine viral diarrhoea virus, and bovine respiratory syncytial virus, with comparison to existing ELISA detection methods

Anderson, Steve; Wakeley, Phil; Wibberley, G.; Webster, Kath; Sawyer, Jason

doi:10.1016/j.jim.2011.01.010

Cited by 41 publications

(32 citation statements)

References 19 publications

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“…In the Luminex study, chemical conjugation of the antigens to Luminex beads may have affected the antigen and led to poorer assay performance. 1 The improved results observed in the current study may be because, in the MSD platform, the Multiplex ELISA for detection of bovine respiratory diseases …”

mentioning

confidence: 67%

“…In broad terms, these results are promising. They are better than those observed when using the same antigens in the Luminex multiplex platform, 1 where the performance of the BVDV and BRSV components of a Luminex multiplex assay were significantly worse. In the Luminex study, chemical conjugation of the antigens to Luminex beads may have affected the antigen and led to poorer assay performance.…”

mentioning

confidence: 70%

“…In the Luminex study, chemical conjugation of the antigens to Luminex beads may have affected the antigen and led to poorer assay performance. 1 The improved results observed in the current study may be because, in the MSD platform, the antigens are adsorbed onto the carbon electrodes, analogous to adsorption in standard ELISA plates. As a result, the antigens behave in a functionally more similar manner.…”

mentioning

confidence: 76%

“…These antigens were more concentrated than those used for coating the single-plex ELISA plate wells. 1 The printed plates were shipped to the AHVLA laboratory in a dry state for further use.…”

mentioning

confidence: 99%

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Development and evaluation of a multiplex enzyme-linked immunosorbent assay for the detection of antibodies to bovine respiratory diseases on the Meso Scale Discovery platform

Mayers

Sawyer

2012

J VET Diagn Invest

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Abstract.A multiplex enzyme-linked immunosorbent assay (ELISA) to detect antibodies to Bovine viral diarrhea virus (BVDV), Bovine herpesvirus 1 (BHV-1), Bovine parainfluenza virus 3 (BPV-3), and Bovine respiratory syncytial virus (BRSV) was developed on the commercially available Meso Scale Discovery (MSD) platform. The multiplex ELISA used the same antigens (relatively crude lysates of cultured virus) that are used in the current in-house separate ELISAs. Testing a panel of field samples demonstrated the multiplex assay could simultaneously detect the presence of several antibodies in a single assay and that there was good agreement between the multiplex ELISA and the in-house ELISAs. The use of a multiplex ELISA could offer significant benefits for routine serological testing through reduced staff time, reduced reagent cost, and streamlined laboratory operation.Key words: Bovine respiratory disease; Meso Scale Discovery; multiplex enzyme-linked immunosorbent assay; serology. Mayers and SawyerThe MSD platform combines electrochemiluminescence (ECL) and microarray-style spatial gridding technology to allow the monitoring and detection of up to 10 separate assays within a single well of a 96-well microtiter plate. 5 The MSD 96-well plates are fabricated to contain separate carbon electrodes (from 1 to 10) within each well, which can be coated with different biological molecules. For the current study, the electrodes were coated with the different antigens used in the individual ELISAs. The fundamental basis of the assay is the same as a typical serology ELISA in that a series of incubation and wash steps in 96-well plate format result in the capture of antibodies in a serum sample and subsequent detection by addition of a labeled anti-species secondary antibody. In this case, the label is a proprietary MSD electrochemiluminescent molecule, SULFO-Tag™.a Generation and measurement of the electrochemiluminescent signal are achieved through electrical stimulation of the SULFO-Tag a -labeled antibodies, where only labels in close proximity to the carbon electrodes are detected. Measurement of the resulting light signal from around the individual electrodes occurs in the MSD Sector Imager instrument, which effectively performs the same function as a microtiter plate reader. 8Initial assay development and optimization of parameters such as antigen concentration, serum dilution, and washing conditions were carried out at the AHVLA laboratories in a single-plex format on the MSD platform, where the wells of the MSD 96-well plates contained a single carbon electrode. This provided the opportunity to optimize each assay individually before combining in a multiplex format.Multiplex plates, where different antigens were adsorbed onto the individual carbon electrodes within each well, were printed by MSD using proprietary robotic printing systems. In the current study, 7-plex plates were printed, which consisted of a carbon electrode coated in each of the 4 positive antigens, and 3 electrodes coated in negative antigen (lysate...

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confidence: 67%

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confidence: 70%

“…In the Luminex study, chemical conjugation of the antigens to Luminex beads may have affected the antigen and led to poorer assay performance. 1 The improved results observed in the current study may be because, in the MSD platform, the antigens are adsorbed onto the carbon electrodes, analogous to adsorption in standard ELISA plates. As a result, the antigens behave in a functionally more similar manner.…”

mentioning

confidence: 76%

mentioning

confidence: 99%

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Development and evaluation of a multiplex enzyme-linked immunosorbent assay for the detection of antibodies to bovine respiratory diseases on the Meso Scale Discovery platform

Mayers

Sawyer

2012

J VET Diagn Invest

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“…Recently, other high-throughput multiplexed assays based on the Luminex platform have been developed to detect antibodies against a variety of viral pathogens (32,42,43). Luminex-based assays have shown greater sensitivity and better performance than ELISA with fast turnaround time and high throughput (31).…”

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confidence: 99%

Development of Whole-Virus Multiplex Luminex-Based Serological Assays for Diagnosis of Infections with Kaposi's Sarcoma-Associated Herpesvirus/Human Herpesvirus 8 Homologs in Macaques

Ryan¹,

Rose

2013

Clin. Vaccine Immunol.

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bKaposi's sarcoma-associated herpesvirus (KSHV)/human herpesvirus 8 is a tumorigenic rhadinovirus that is associated with all forms of Kaposi's sarcoma. Current serological detection of KSHV is based on enzyme-linked immunosorbent or immunofluorescence assays that suffer from a variety of problems, including the lack of defined standards for test comparison. While KSHV is the only known human rhadinovirus, two lineages of KSHV-like rhadinoviruses are found in Old World primates: the RV1 lineage includes KSHV and retroperitoneal fibromatosis herpesvirus (RFHV) in macaques, and the RV2 lineage includes RRV and MneRV2 from different macaque species. To develop animal models of KSHV-associated diseases, we developed quantitative multiplex bead-based serological assays to detect antibodies against rhadinovirus antigens. Proteins from KSHV (RV1) and MneRV2 (RV2) virions were coupled to spectrally distinct fluorescent beads and used in Luminex flow cytometry-based assays to detect immune responses in macaques. Both assays showed large dynamic ranges with high levels of seroreactivity to both KSHV and MneRV2 proteins. A large set of macaque serum samples from the Washington National Primate Research Center was screened, and most of the samples (82%) were positive in both assays, consistent with the high level of RV1-RV2 coinfection detected by PCR. The macaque sera showed broad, variable, and unique serological responses to the different viral antigens, allowing an initial seroprevalence to be determined for the macaque viruses. The Luminex assays offer a novel multiplexed approach to assess rhadinovirus infection patterns in both humans and nonhuman primates. This will help advance our understanding of rhadinovirus biology and associated host immunological responses.

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Immunoassay

Taylor¹

2016

Kirk-Othmer Encyclopedia of Chemical Technology

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Immunoassay, a diagnostic and measurement method that utilizes antibodies to detect and quantify specific analytes (antigens), is characterized by high analyte specificity, high sensitivity (routinely detecting ng to pg analyte levels), and rapid data output. It is amenable to a variety of formats, from single immunoassays carried out manually, to fully automated instruments capable of performing multiple assays on the same or different analytes. Immunoassay quantification methods are based on immunoprecipitation, such as agglutination assays, or on specific indicator reagents linked to antibody or antigen reagents used in the immunoassay. These include radioisotopes for radioimmunoassay, enzymes for enzyme‐linked immunoassay, fluorophores for fluorescence immunoassay, and chemicals or proteins for chemiluminescent or bioluminescent immunoassay. Immunoassay technology is continuing to evolve and applications have moved beyond traditional clinical settings to industry and the environment. Immunoassays are also serving as the basis for newer measurement technologies including hybrid immunoassay‐nucleic acid probe assays, immunobiosensors, and antibody‐based microarrays.

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Development and evaluation of a Luminex multiplex serology assay to detect antibodies to bovine herpes virus 1, parainfluenza 3 virus, bovine viral diarrhoea virus, and bovine respiratory syncytial virus, with comparison to existing ELISA detection methods

Cited by 41 publications

References 19 publications

Development and evaluation of a multiplex enzyme-linked immunosorbent assay for the detection of antibodies to bovine respiratory diseases on the Meso Scale Discovery platform

Development and evaluation of a multiplex enzyme-linked immunosorbent assay for the detection of antibodies to bovine respiratory diseases on the Meso Scale Discovery platform

Development of Whole-Virus Multiplex Luminex-Based Serological Assays for Diagnosis of Infections with Kaposi's Sarcoma-Associated Herpesvirus/Human Herpesvirus 8 Homologs in Macaques

Immunoassay

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