Development of a lab-on-a-chip device for diagnosis of plant pathogens

Julich, Sandra; Riedel, Marko; Kielpinski, Mark; Urban, Matthias; Kretschmer, Robert; Wagner, Ştefan; Fritzsche, Wolfgang; Henkel, Thomas; Möller, Robert; Werres, Sabine

doi:10.1016/j.bios.2011.03.035

Cited by 59 publications

(38 citation statements)

References 42 publications

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“…[13] Compared to chip-based methods [14][15][16][17][18][19][20][21][22] the use of magnetic particles benefits from improved hybridization kinetics. By employing biotin-labeled primers for the amplification of target DNA with polymerase chain reaction (PCR), the PCR products can bind directly onto the magnetic beads.…”

Section: Magnetic Particlesmentioning

confidence: 99%

Discrimination of clostridium species using a magnetic bead based hybridization assay

Pahlow

Seise

Pollok

et al. 2014

Biophotonics: Photonic Solutions for Better Health Care IV

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Clostridium chauvoei is the causative agent of blackleg, which is an endogenous bacterial infection. Mainly cattle and other ruminants are affected. The symptoms of blackleg are very similar to those of malignant edema, an infection caused by Clostridium septicum. [1, 2] Therefore a reliable differentiation of Clostridium chauvoei from other Clostridium species is required. Traditional microbiological detection methods are time consuming and laborious. Additionally, the unique identification is hindered by the overgrowing tendency of swarming Clostridium septicum colonies when both species are present. [1, 3, 4] Thus, there is a crucial need to improve and simplify the specific detection of Clostridium chauvoei and Clostridium septicum.Here we present an easy and fast Clostridium species discrimination method combining magnetic beads and fluorescence spectroscopy. Functionalized magnetic particles exhibit plentiful advantages, like their simple manipulation in combination with a large binding capacity of biomolecules. A specific region of the pathogenic DNA is amplified and labelled with biotin by polymerase chain reaction (PCR). These PCR products were then immobilized on magnetic beads exploiting the strong biotin-streptavidin interaction. The specific detection of different Clostridium species is achieved by using fluorescence dye labeled probe DNA for the hybridization with the immobilized PCR products. Finally, the samples were investigated by fluorescence spectroscopy. [5]

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Section: Magnetic Particlesmentioning

confidence: 99%

Discrimination of clostridium species using a magnetic bead based hybridization assay

Pahlow

Seise

Pollok

et al. 2014

Biophotonics: Photonic Solutions for Better Health Care IV

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“…Saliva samples from participants were directly used without any purification process, showing the potential of point-of-care SNP genotyping. A few more examples include the identification of plant pathogens [86], Escherichia coli ( E. coli ) and hepatitis B virus [87], human disease of non-syndromic sensorineural hearing loss (NSSNHL) [88], and many others.…”

Section: Miniaturizing Platforms Of Snp Genotypingmentioning

confidence: 99%

Efficient SNP Discovery by Combining Microarray and Lab-on-a-Chip Data for Animal Breeding and Selection

et al. 2015

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The genetic markers associated with economic traits have been widely explored for animal breeding. Among these markers, single-nucleotide polymorphism (SNPs) are gradually becoming a prevalent and effective evaluation tool. Since SNPs only focus on the genetic sequences of interest, it thereby reduces the evaluation time and cost. Compared to traditional approaches, SNP genotyping techniques incorporate informative genetic background, improve the breeding prediction accuracy and acquiesce breeding quality on the farm. This article therefore reviews the typical procedures of animal breeding using SNPs and the current status of related techniques. The associated SNP information and genotyping techniques, including microarray and Lab-on-a-Chip based platforms, along with their potential are highlighted. Examples in pig and poultry with different SNP loci linked to high economic trait values are given. The recommendations for utilizing SNP genotyping in nimal breeding are summarized.

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“…However, these systems do not employ real-time detection, and thus require additional methods or devices such as gel electrophoresis for the quantification of amplicons. In recent years, several real-time microchip PCR systems integrated with optical detection systems have been developed with promising results, but mostly for clinical diagnosis applications [13]–[22], Julich et al has reported the one real-time microchip PCR system for plant pathogen detection [23], but the system still requires various auxiliary instruments such as a laptop, fluidic pump, electrical readout and temperature regulation system which are larger than the laptop size.…”

Section: Introductionmentioning

confidence: 99%

Development of a Real-Time Microchip PCR System for Portable Plant Disease Diagnosis

et al. 2013

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Rapid and accurate detection of plant pathogens in the field is crucial to prevent the proliferation of infected crops. Polymerase chain reaction (PCR) process is the most reliable and accepted method for plant pathogen diagnosis, however current conventional PCR machines are not portable and require additional post-processing steps to detect the amplified DNA (amplicon) of pathogens. Real-time PCR can directly quantify the amplicon during the DNA amplification without the need for post processing, thus more suitable for field operations, however still takes time and require large instruments that are costly and not portable. Microchip PCR systems have emerged in the past decade to miniaturize conventional PCR systems and to reduce operation time and cost. Real-time microchip PCR systems have also emerged, but unfortunately all reported portable real-time microchip PCR systems require various auxiliary instruments. Here we present a stand-alone real-time microchip PCR system composed of a PCR reaction chamber microchip with integrated thin-film heater, a compact fluorescence detector to detect amplified DNA, a microcontroller to control the entire thermocycling operation with data acquisition capability, and a battery. The entire system is 25×16×8 cm3 in size and 843 g in weight. The disposable microchip requires only 8-µl sample volume and a single PCR run consumes 110 mAh of power. A DNA extraction protocol, notably without the use of liquid nitrogen, chemicals, and other large lab equipment, was developed for field operations. The developed real-time microchip PCR system and the DNA extraction protocol were used to successfully detect six different fungal and bacterial plant pathogens with 100% success rate to a detection limit of 5 ng/8 µl sample.

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Development of a lab-on-a-chip device for diagnosis of plant pathogens

Cited by 59 publications

References 42 publications

Discrimination of clostridium species using a magnetic bead based hybridization assay

Discrimination of clostridium species using a magnetic bead based hybridization assay

Efficient SNP Discovery by Combining Microarray and Lab-on-a-Chip Data for Animal Breeding and Selection

Development of a Real-Time Microchip PCR System for Portable Plant Disease Diagnosis

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