scholarly journals Preserved Microarrays for Simultaneous Detection and Identification of Six Fungal Potato Pathogens with the Use of Real-Time PCR in Matrix Format

Biosensors ◽  
2018 ◽  
Vol 8 (4) ◽  
pp. 129 ◽  
Author(s):  
Maksim Nikitin ◽  
Ksenia Deych ◽  
Inessa Grevtseva ◽  
Natalya Girsova ◽  
Maria Kuznetsova ◽  
...  
Keyword(s):  
Detection Limit ◽  
Plant Pathogens ◽  
Simultaneous Detection ◽  
Morphological Identification ◽  
Target Species ◽  
Test Systems ◽  
Detection And Identification ◽  
Freeze Dried ◽  
Field Samples ◽  
Potato Pathogens

Fungal diseases of plants are of great economic importance causing 70–80% of crop losses associated with microbial plant pathogens. Advanced on-site disease diagnostics is very important to maximize crop productivity. In this study, diagnostic systems have been developed for simultaneous detection and identification of six fungal pathogens using 48-well microarrays (micromatrices) for qPCR. All oligonucleotide sets were tested for their specificity using 59 strains of target and non-target species. Detection limit of the developed test systems varied from 0.6 to 43.5 pg of DNA depending on target species with reproducibility within 0.3−0.7% (standard deviation). Diagnostic efficiency of test systems with stabilized and freeze-dried PCR master-mixes did not significantly differ from that of freshly prepared microarrays, though detection limit increased. Validation of test systems on 30 field samples of potato plants showed perfect correspondence with the results of morphological identification of pathogens. Due to the simplicity of the analysis and the automated data interpretation, the developed microarrays have good potential for on-site use by technician-level personnel, as well as for high-throughput monitoring of fungal potato pathogens.

scholarly journals A Macroarray System for the Detection of Fungal and Oomycete Pathogens of Solanaceous Crops

Plant Disease ◽  
2008 ◽  
Vol 92 (6) ◽  
pp. 953-960 ◽  
Author(s):  
Ning Zhang ◽  
Meaghan L. McCarthy ◽  
Christine D. Smart
Keyword(s):  
Pathogen Detection ◽  
Detection System ◽  
Simultaneous Detection ◽  
Rrna Genes ◽  
Identification System ◽  
Nylon Membrane ◽  
Detection And Identification ◽  
Field Samples ◽  
Nontarget Species ◽  
Plant Disease Management

There are numerous fungal and oomycete pathogens that cause severe damage to solanaceous crops. Rapid and accurate detection and identification of these pathogens is critical for plant disease management. Recently, DNA array technology has been successfully applied for simultaneous detection of multiple microorganisms from various habitats. The goal of this project was to develop a multiplex detection and identification system for major fungal and oomycete pathogens of solanaceous crops. To facilitate this goal, we used a membrane-based macroarray technology that included at least two specific oligonucleotides per pathogen. Based on the internal transcribed spacer sequences of the rRNA genes, 105 oligonucleotides (17 to 27 bases long) specific for 25 pathogens of solanaceous crops were designed and spotted on a nylon membrane. The array was tested against the 25 target pathogen species, 46 infected field samples, and a number of nontarget species. Our results indicate that the oligonucleotide-based macroarray detection system is a reliable and effective method for pathogen detection and identification even when multiple pathogens are present in a field sample.

scholarly journals A New Multiplex Real-Time RT-PCR for Simultaneous Detection and Differentiation of Avian Bornaviruses

Viruses ◽  
2021 ◽  
Vol 13 (7) ◽  
pp. 1358
Author(s):  
Brigitte Sigrist ◽  
Jessica Geers ◽  
Sarah Albini ◽  
Dennis Rubbenstroth ◽  
Nina Wolfrum
Keyword(s):  
Real Time ◽  
Simultaneous Detection ◽  
Epidemiological Studies ◽  
Virus Species ◽  
Rt Pcr ◽  
P Gene ◽  
Field Samples ◽  
Causative Agents ◽  
Species Specific ◽  
Proventricular Dilatation Disease

Avian bornaviruses were first described in 2008 as the causative agents of proventricular dilatation disease (PDD) in parrots and their relatives (Psittaciformes). To date, 15 genetically highly diverse avian bornaviruses covering at least five viral species have been discovered in different bird orders. Currently, the primary diagnostic tool is the detection of viral RNA by conventional or real-time RT-PCR (rRT-PCR). One of the drawbacks of this is the usage of either specific assays, allowing the detection of one particular virus, or of assays with a broad detection spectrum, which, however, do not allow for the simultaneous specification of the detected virus. To facilitate the simultaneous detection and specification of avian bornaviruses, a multiplex real-time RT-PCR assay was developed. Whole-genome sequences of various bornaviruses were aligned. Primers were designed to recognize conserved regions within the overlapping X/P gene and probes were selected to detect virus species-specific regions within the target region. The optimization of the assay resulted in the sensitive and specific detection of bornaviruses of Psittaciformes, Passeriformes, and aquatic birds. Finally, the new rRT-PCR was successfully employed to detect avian bornaviruses in field samples from various avian species. This assay will serve as powerful tool in epidemiological studies and will improve avian bornavirus detection.

ANALYTICAL SYSTEMS BASED ON RECOMBINASE POLYMERASE AMPLIFICATION FOR RAPID DETECTION OF VIRAL, VIROID AND BACTERIAL PLANT PATHOGENS

2021 ◽  
Vol 1 (19) ◽  
pp. 242-244
Author(s):  
A.V. Ivanov ◽  
A.V. Zherdev ◽  
B.B. Dzantiev
Keyword(s):  
Potato Virus ◽  
Rapid Detection ◽  
Plant Pathogens ◽  
Potato Virus X ◽  
Potato Spindle Tuber Viroid ◽  
Recombinase Polymerase Amplification ◽  
Analysis Time ◽  
Potato Blackleg ◽  
Test Systems ◽  
Dickeya Solani

Test systems have been developed for the detection of phytopathogens, combining recombinase polymerase amplification and membrane test strips. Test systems provide detection of potato virus X, potato spindle tuber viroid, potato blackleg pathogen (Dickeya solani), as well as multi-analysis of three viruses. Amplification is carried out at 37 °C. The analysis time does n ot exceed 30 min.

A new one-step multiplex PCR assay for simultaneous detection and identification of avian haemosporidian parasites

2018 ◽  
Vol 118 (1) ◽  
pp. 191-201 ◽  
Author(s):  
Arif Ciloglu ◽  
Vincenzo A. Ellis ◽  
Rasa Bernotienė ◽  
Gediminas Valkiūnas ◽  
Staffan Bensch
Keyword(s):  
Multiplex Pcr ◽  
Simultaneous Detection ◽  
Pcr Assay ◽  
Haemosporidian Parasites ◽  
Multiplex Pcr Assay ◽  
Detection And Identification ◽  
One Step

Simultaneous detection and identification of multigas pollutants using filament-induced nonlinear spectroscopy

2007 ◽  
Vol 90 (10) ◽  
pp. 101106 ◽  
Author(s):  
H. L. Xu ◽  
Y. Kamali ◽  
C. Marceau ◽  
P. T. Simard ◽  
W. Liu ◽  
...  
Keyword(s):  
Simultaneous Detection ◽  
Nonlinear Spectroscopy ◽  
Detection And Identification
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