A multiplexed PCR-coupled liquid bead array for the simultaneous detection of four biothreat agents

Wilson, W J; Erler, Anne Marie; Nasarabadi, Shanavaz; Skowronski, Evan W.; Imbro, Paula M.

doi:10.1016/j.mcp.2004.10.005

Cited by 63 publications

(59 citation statements)

References 25 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The xMAP technology is based on uniquely color-coded microspheres, which allows as many as one hundred different species to be detected in a single reaction. This technology has been used for the detection of several species of bacteria and fungi (7,8,9,11,13,24,31). Recently, xMAP technology has been used in several diagnostic kits for the detection of bacterial and viral pathogens.…”

mentioning

confidence: 99%

Identification of Genotypically Diverse Cryptococcus neoformans and Cryptococcus gattii Isolates by Luminex xMAP Technology

et al. 2007

View full text Add to dashboard Cite

A Luminex suspension array, which had been developed for identification of Cryptococcus neoformans andCryptococcus gattii isolates, was tested by genotyping a set of 58 mostly clinical isolates. All genotypes of C. neoformans and C. gattii were included. In addition, cerebrospinal fluid (CSF) obtained from patients with cryptococcal meningitis was used to investigate the feasibility of the technique for identification of the infecting strain. The suspension array correctly identified haploid isolates in all cases. Furthermore, hybrid isolates possessing two alleles of the Luminex probe region could be identified as hybrids. In CSF specimens, the genotype of the cryptococcal strains responsible for infection could be identified after optimization of the PCR conditions. However, further optimization of the DNA extraction protocol is needed to enhance the usability of the method in clinical practice.

show abstract

mentioning

confidence: 99%

Identification of Genotypically Diverse Cryptococcus neoformans and Cryptococcus gattii Isolates by Luminex xMAP Technology

et al. 2007

View full text Add to dashboard Cite

show abstract

“…The suspension microarray system has recently been demonstrated to be a rapid, sensitive, and specific assay for detection of bacterial pathogens, including L. monocytogenes (8,25). The Luminex (Austin, TX) suspension array is simply a transfer of the microarray format from a glass slide (planar microarray) to a high-throughput and efficient bead format ("suspension microarray").…”

mentioning

confidence: 99%

Suspension Microarray with Dendrimer Signal Amplification Allows Direct and High-Throughput Subtyping of Listeria monocytogenes from Genomic DNA

Borucki

Reynolds²,

Call

et al. 2005

J Clin Microbiol

View full text Add to dashboard Cite

Listeria monocytogenes is a significant cause of food-borne disease and mortality; therefore, epidemiological investigations of this pathogen require subtyping methods that are rapid, discriminatory, and reproducible. Although conventional microarray subtyping analysis has been shown to be both high resolution and genetically informative, it is still relatively low throughput and technically challenging. Suspension microarray technology eliminates the technical issues associated with planar microarrays and allows high-throughput subtyping of L. monocytogenes strains. In this study, a suspension array assay using dendrimer signal amplification allowed rapid and accurate serovar identification of L. monocytogenes strains using genomic DNA as a target. The ability to subtype genomic DNA without PCR amplification allows probes to be designed for many different regions within the bacterial genome and should allow high-resolution subtyping not possible with multiplex PCR.Listeria monocytogenes is a gram-positive bacterial pathogen that causes significant morbidity and mortality. Human listeriosis may occur as widespread epidemics or, more commonly, as sporadic cases (18). It is likely, however, that a significant percentage of sporadic cases are unrecognized single-source outbreaks (19). Epidemiological investigation of epidemic and sporadic cases of listeriosis requires molecular characterization to allow the identification of specific subtypes.Molecular subtyping techniques have identified two major phylogenetic divisions within the species, with division I including serotypes 1/2b, 3b, and 4b and division II consisting of serotypes 1/2a, 1/2c, 3a, and 3c (15,16). A third division consisting of serotypes 4a, 4c, and a subset of 4b strains has also been described (4, 24). Although 13 serotypes of L. monocytogenes have been described (20), only three serotypes (1/2a, 1/2b, and 4b) are responsible for the vast majority of clinical cases (23).L. monocytogenes subtypes are usually characterized by serotyping and then further subtyped using pulsed-field gel electrophoresis (10) or ribotyping. Due to the importance of L. monocytogenes epidemiology to human health, new subtyping technologies are constantly being developed in the hope of increasing the resolution, speed, and reproducibility of L. monocytogenes subtyping. The most recent examples of this are multilocus sequence subtyping (12, 17) and microarray genomic analysis (1,5,26). Recent studies have shown that a relatively simple genotyping microarray has subtyping resolution comparable to that of pulsed-field gel electrophoresis (AscI and ApaI digestion) and superior to that of multilocus sequence subtyping (six housekeeping genes) and ribotyping (3). L. monocytogenes subtyping using DNA microarrays is also genetically informative because it can identify the genes or alleles that characterize subtypes.DNA microarrays are typically composed of DNA "probes" (nucleic acids of known sequence) that are bound to a solid substrate, such as a glass microarray slide (referred...

show abstract

“…Out of 56 articles on the instrument limit of detection, 17 articles were on real-time PCR (6,7,11,23,25,27,39,41,45,49,51,53,56,58,60,70,72), 6 were on PCR (13,31,48,57,76) (37,55,78), and 4 were on mass spectrometry (8, 9, 28, 43) (Fig. 1).…”

Section: Resultsmentioning

confidence: 99%

Implications of Limits of Detection of Various Methods for Bacillus anthracis in Computing Risks to Human Health

Herzog

McLennan

Pandey

et al. 2009

Appl Environ Microbiol

View full text Add to dashboard Cite

Used for decades for biological warfare, Bacillus anthracis (category A agent) has proven to be highly stable and lethal. Quantitative risk assessment modeling requires descriptive statistics of the limit of detection to assist in defining the exposure. Furthermore, the sensitivities of various detection methods in environmental matrices are vital information for first responders. A literature review of peer-reviewed journal articles related to methods for detection of B. anthracis was undertaken. Articles focused on the development or evaluation of various detection approaches, such as PCR, real-time PCR, immunoassay, etc. Real-time PCR and PCR were the most sensitive methods for the detection of B. anthracis, with median instrument limits of detection of 430 and 440 cells/ml, respectively. There were very few peer-reviewed articles on the detection methods for B. anthracis in the environment. The most sensitive limits of detection for the environmental samples were 0.1 CFU/g for soil using PCR-enzyme-linked immunosorbent assay (ELISA), 17 CFU/liter for air using an ELISA-biochip system, 1 CFU/liter for water using cultivation, and 1 CFU/cm 2 for stainless steel fomites using cultivation. An exponential dose-response model for the inhalation of B. anthracis estimates of risk at concentrations equal to the environmental limit of detection determined the probability of death if untreated to be as high as 0.520. Though more data on the environmental limit of detection would improve the assumptions made for the risk assessment, this study's quantification of the risk posed by current limitations in the knowledge of detection methods should be considered when employing those methods in environmental monitoring and cleanup strategies.

show abstract

A multiplexed PCR-coupled liquid bead array for the simultaneous detection of four biothreat agents

Cited by 63 publications

References 25 publications

Identification of Genotypically Diverse Cryptococcus neoformans and Cryptococcus gattii Isolates by Luminex xMAP Technology

Identification of Genotypically Diverse Cryptococcus neoformans and Cryptococcus gattii Isolates by Luminex xMAP Technology

Suspension Microarray with Dendrimer Signal Amplification Allows Direct and High-Throughput Subtyping of Listeria monocytogenes from Genomic DNA

Implications of Limits of Detection of Various Methods for Bacillus anthracis in Computing Risks to Human Health

Contact Info

Product

Resources

About