Pathogen Detection Using a Liquid Array Technology*

Battaglia, Amanda; Schweighardt, Andrew J.; Wallace, Margaret M.

doi:10.1111/j.1556-4029.2011.01708.x

Cited by 13 publications

(8 citation statements)

References 10 publications

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“…Alternative approaches to parallel detection tests include melting curve analysis [37] and bead-based Luminex assays [38], [39]. However, while rapid, highly specific and reasonably sensitive, the parallelity of these technologies is practically limited to about ten different species in a single test.…”

Section: Discussionmentioning

confidence: 99%

A Novel Rapid DNA Microarray Assay Enables Identification of 37 Mycoplasma Species and Highlights Multiple Mycoplasma Infections

et al. 2012

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Mycoplasmas comprise a conglomerate of pathogens and commensals occurring in humans and animals. The genus Mycoplasma alone contains more than 120 species at present, and new members are continuously being discovered. Therefore, it seems promising to use a single highly parallel detection assay rather than develop separate tests for each individual species. In this study, we have designed a DNA microarray carrying 70 oligonucleotide probes derived from the 23S rRNA gene and 86 probes from the tuf gene target regions. Following a PCR amplification and biotinylation step, hybridization on the array was shown to specifically identify 31 Mycoplasma spp., as well as 3 Acholeplasma spp. and 3 Ureaplasma spp. Members of the Mycoplasma mycoides cluster can be recognized at subgroup level. This procedure enables parallel detection of Mollicutes spp. occurring in humans, animals or cell culture, from mono- and multiple infections, in a single run. The main advantages of the microarray assay include ease of operation, rapidity, high information content, and affordability. The new test's analytical sensitivity is equivalent to that of real-time PCR and allows examination of field samples without the need for culture. When 60 field samples from ruminants and birds previously analyzed by denaturing-gradient gel electrophoresis (DGGE) were tested by the microarray assay both tests identified the same agent in 98.3% of the cases. Notably, microarray testing revealed an unexpectedly high proportion (35%) of multiple mycoplasma infections, i.e., substantially more than DGGE (15%). Two of the samples were found to contain four different Mycoplasma spp. This phenomenon deserves more attention, particularly its implications for epidemiology and treatment.

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Section: Discussionmentioning

confidence: 99%

A Novel Rapid DNA Microarray Assay Enables Identification of 37 Mycoplasma Species and Highlights Multiple Mycoplasma Infections

et al. 2012

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“…A blue fitting curve has been derived as, where x is the concentration of E. coli and y is the imaginary impedance peak, R et. (8) Then we performed a measurement of prepared bacteria solution with calculated concentration of 333 cells per milliliter. The result is shown as the thick orange curve in Fig 5 (b), where R et equals 25727.4 Ohm which is plotted as the green cross in Fig 5 (c).…”

Section: Smartphone Software Applicationmentioning

confidence: 99%

“…However, fluorescence detection needs dye labeling [7] which requires high expense and professional training for operation. DNA-biosensor is also required to perform Polymerase chain reaction (PCR) as the first step [8] which is a complicated procedure in need of complicated facilities. So researchers also invented a series of label-free detection methods including quartz crystal microbalance (QCM) [9] , microfluidic [10] and electrochemical methods [3] .…”

Section: Introductionmentioning

confidence: 99%

Smartphone based portable bacteria pre-concentrating microfluidic sensor and impedance sensing system

Jiang

Wang

Chao

et al. 2014

Sensors and Actuators B: Chemical

148

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“…The procedure requires some hands-on time and additional preanalytic effort. Experiences in direct pathogen detection are rather sparse and the overall sensitivity seems to be insufficient for direct trace detection of low-copy microbial DNA/cell counts [64]. However, the approach is quite smart for multiplexing applications and may be applied more routinely in the future.…”

Section: Flow Cytometrymentioning

confidence: 99%

Nucleic Acid Amplification Techniques

Lehmann¹,

Schmitz

2015

Modern Techniques for Pathogen Detection

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Nucleic acid amplification techniques (NATs), more commonly but imprecisely called polymerase chain reaction (PCR), have evolved, among others, as central tools for pathogen identification and strain typing. The huge field of applications of the technique in all its modifications already marks the centerpiece of diagnostic approaches and has not only opened the door to a completely new understanding of elementary and sophisticated metabolic processes but also is the basis of the most important trends in life science industries. In a sense, it acts as the lens of a molecular microscope which visualizes the code from the genomic book, while its meanings have to be deciphered by a further armory of laboratory avenues.In 1983, PCR was invented by Kary Mullis, for which he later (1993) was awarded with the Nobel Prize in Chemistry along with Michael Smith [1]. The protocol is based on targeting specific DNA or rDNA sequences with more or less specific short tracer sequences (primers/oligonucleotides, colloquially oligos) binding to flanking regions of the core sequence to be proliferated. A succession of thermal incubation steps defined by length and temperature are cycled and conduct double-stranded DNA (dsDNA) melting, that is, separation of the target DNA into antiparallel, complementary, single-stranded DNA (ssDNA) strands, specific binding of primers (annealing), and enzymatic amplification out of the basic set of four nucleotide monomers of the sequence embedded. In continuation, the newly synthesized dsDNA copies, selected in length by the specific primer binding sites (PBSs), themselves serve as templates for further replication rounds, setting in motion a chain reaction in which the DNA template is exponentially amplified. The core enzyme is a DNA polymerase, which was heat-sensitive at time of invention but subsequently was destroyed at the melting temperature. To gain useful numbers of target copies, an expensive and time-consuming replacement of the enzyme was needed at the beginning of each of the usually 20-40 replication cycles. The discovery of an extraordinarily thermostable DNA polymerase purified from the thermophilic bacterium Thermophilus aquaticus, which naturally lives in hot (50-80 ∘ C) environments such as hot springs [2], Modern Techniques for Pathogen Detection, First Edition. Edited by Jürgen Popp and Michael Bauer.

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Pathogen Detection Using a Liquid Array Technology*

Cited by 13 publications

References 10 publications

A Novel Rapid DNA Microarray Assay Enables Identification of 37 Mycoplasma Species and Highlights Multiple Mycoplasma Infections

A Novel Rapid DNA Microarray Assay Enables Identification of 37 Mycoplasma Species and Highlights Multiple Mycoplasma Infections

Smartphone based portable bacteria pre-concentrating microfluidic sensor and impedance sensing system

Nucleic Acid Amplification Techniques

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