Multiplex Detection of Plant Pathogens Using a Microsphere Immunoassay Technology
Publication Date
April 26, 2013
Journal
PLOS ONE
Authors
Ratthaphol Charlermroj, Orawan Himananto, Channarong Seepiban, Mallika Kumpoosiri, et al
Volume
8
Issue
4
Pages
e62344
DOI
https://dx.plos.org/10.1371/journal.pone.0062344
Publisher URL
http://journals.plos.org/plosone/article?id=10.1371%2Fjournal.pone.0062344
PubMed
http://www.ncbi.nlm.nih.gov/pubmed/23638044
PubMed Central
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3637204
Europe PMC
http://europepmc.org/abstract/MED/23638044
Web of Science
000318343700050
Scopus
84876836450
Mendeley
http://www.mendeley.com/research/multiplex-detection-plant-pathogens-using-microsphere-immunoassay-technology
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Mendeley | Further Information

{"title"=>"Multiplex Detection of Plant Pathogens Using a Microsphere Immunoassay Technology", "type"=>"journal", "authors"=>[{"first_name"=>"Ratthaphol", "last_name"=>"Charlermroj", "scopus_author_id"=>"25026853500"}, {"first_name"=>"Orawan", "last_name"=>"Himananto", "scopus_author_id"=>"36622702200"}, {"first_name"=>"Channarong", "last_name"=>"Seepiban", "scopus_author_id"=>"36626344500"}, {"first_name"=>"Mallika", "last_name"=>"Kumpoosiri", "scopus_author_id"=>"47561330500"}, {"first_name"=>"Nuchnard", "last_name"=>"Warin", "scopus_author_id"=>"23111597300"}, {"first_name"=>"Michalina", "last_name"=>"Oplatowska", "scopus_author_id"=>"37561732700"}, {"first_name"=>"Oraprapai", "last_name"=>"Gajanandana", "scopus_author_id"=>"23110409300"}, {"first_name"=>"Irene R.", "last_name"=>"Grant", "scopus_author_id"=>"7203073137"}, {"first_name"=>"Nitsara", "last_name"=>"Karoonuthaisiri", "scopus_author_id"=>"24765223200"}, {"first_name"=>"Christopher T.", "last_name"=>"Elliott", "scopus_author_id"=>"7202077903"}], "year"=>2013, "source"=>"PLoS ONE", "identifiers"=>{"issn"=>"19326203", "pui"=>"368819800", "doi"=>"10.1371/journal.pone.0062344", "sgr"=>"84876836450", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "pmid"=>"23638044", "scopus"=>"2-s2.0-84876836450"}, "id"=>"cf558530-9496-3e1c-9be1-17fa7f705cc3", "abstract"=>"Plant pathogens are a serious problem for seed export, plant disease control and plant quarantine. Rapid and accurate screening tests are urgently required to protect and prevent plant diseases spreading worldwide. A novel multiplex detection method was developed based on microsphere immunoassays to simultaneously detect four important plant pathogens: a fruit blotch bacterium Acidovorax avenae subsp. citrulli (Aac), chilli vein-banding mottle virus (CVbMV, potyvirus), watermelon silver mottle virus (WSMoV, tospovirus serogroup IV) and melon yellow spot virus (MYSV, tospovirus). An antibody for each plant pathogen was linked on a fluorescence-coded magnetic microsphere set which was used to capture corresponding pathogen. The presence of pathogens was detected by R-phycoerythrin (RPE)-labeled antibodies specific to the pathogens. The assay conditions were optimized by identifying appropriate antibody pairs, blocking buffer, concentration of RPE-labeled antibodies and assay time. Once conditions were optimized, the assay was able to detect all four plant pathogens precisely and accurately with substantially higher sensitivity than enzyme-linked immunosorbent assay (ELISA) when spiked in buffer and in healthy watermelon leaf extract. The assay time of the microsphere immunoassay (1 hour) was much shorter than that of ELISA (4 hours). This system was also shown to be capable of detecting the pathogens in naturally infected plant samples and is a major advancement in plant pathogen detection.", "link"=>"http://www.mendeley.com/research/multiplex-detection-plant-pathogens-using-microsphere-immunoassay-technology", "reader_count"=>38, "reader_count_by_academic_status"=>{"Unspecified"=>3, "Professor > Associate Professor"=>1, "Researcher"=>10, "Student > Ph. D. Student"=>12, "Student > Postgraduate"=>1, "Student > Master"=>8, "Student > Bachelor"=>2, "Lecturer"=>1}, "reader_count_by_user_role"=>{"Unspecified"=>3, "Professor > Associate Professor"=>1, "Researcher"=>10, "Student > Ph. D. Student"=>12, "Student > Postgraduate"=>1, "Student > Master"=>8, "Student > Bachelor"=>2, "Lecturer"=>1}, "reader_count_by_subject_area"=>{"Engineering"=>1, "Unspecified"=>4, "Environmental Science"=>1, "Biochemistry, Genetics and Molecular Biology"=>3, "Mathematics"=>1, "Agricultural and Biological Sciences"=>23, "Medicine and Dentistry"=>2, "Chemistry"=>1, "Immunology and Microbiology"=>1, "Economics, Econometrics and Finance"=>1}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>1}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>2}, "Chemistry"=>{"Chemistry"=>1}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>1}, "Economics, Econometrics and Finance"=>{"Economics, Econometrics and Finance"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>23}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>3}, "Mathematics"=>{"Mathematics"=>1}, "Unspecified"=>{"Unspecified"=>4}, "Environmental Science"=>{"Environmental Science"=>1}}, "reader_count_by_country"=>{"Burkina Faso"=>1, "United States"=>1, "Poland"=>1, "United Kingdom"=>1, "Portugal"=>1, "Spain"=>1}, "group_count"=>2}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1044812"], "description"=>"<p>A mixture of antibody-coated microsphere, MPC react to <i>Acidovorax avenae</i> subsp. <i>citrulli</i> (Aac), 1B4 specific to chilli vein-banding mottle virus (CVbMV), 2D6 specific to watermelon silver mottle virus (WSMoV) and 5E7 specific to melon yellow spot virus (MYSV), was tested with a single antigen and no pathogen using (A) 1% skimmed milk, (B) 1% casein or (C) 1% bovine serum albumin (BSA) as the blocking agent. Mixture of RPE-labeled antibodies, MPC, 1G8, A3 and 5E7, were used as a detecting system for Aac, CVbMV, WSMoV and MYSV, respectively. Y-axis is median fluorescent intensity (MFI). Each dataset was plotted as a mean of duplicates ± standard deviation.</p>", "links"=>[], "tags"=>["Agricultural biotechnology", "crops", "Crop diseases", "Pest control", "Biochemistry", "immunochemistry", "biotechnology", "immunology", "Immunologic techniques", "immunoassays", "Plant science", "Plant pathology", "Plant pathogens", "blocking"], "article_id"=>692544, "categories"=>["Medicine", "Biological Sciences"], "users"=>["Ratthaphol Charlermroj", "Orawan Himananto", "Channarong Seepiban", "Mallika Kumpoosiri", "Nuchnard Warin", "Michalina Oplatowska", "Oraprapai Gajanandana", "Irene R. Grant", "Nitsara Karoonuthaisiri", "Christopher T. Elliott"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0062344.g003", "stats"=>{"downloads"=>0, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Selection_of_blocking_buffers_/692544", "title"=>"Selection of blocking buffers.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-04-26 00:42:24"}
  • {"files"=>["https://ndownloader.figshare.com/files/1044809"], "description"=>"<p>The detection of (A) <i>Acidovorax avenae</i> subsp. <i>citrulli</i> (Aac) and (B) no antigen using eight antibodies-coated microsphere and R-Phycoerythrin (RPE) labeled antibodies, including 11E5 antibody. The detection of (C) Aac, (D) chilli vein-banding mottle virus (CVbMV), (E) watermelon silver mottle virus (WSMoV), (F) melon yellow spot virus (MYSV) detection and (G) no antigen with seven antibodies- coated microsphere and RPE-labeled antibodies without using 11E5 antibody. X-axis is antibody-coated microsphere and y-axis is median fluorescent intensity (MFI) from each RPE-labeled antibody. (H) Summary of selected antibody pair sets for the detection of the four plant pathogens.</p>", "links"=>[], "tags"=>["Agricultural biotechnology", "crops", "Crop diseases", "Pest control", "Biochemistry", "immunochemistry", "biotechnology", "immunology", "Immunologic techniques", "immunoassays", "Plant science", "Plant pathology", "Plant pathogens", "antibody", "pairs", "multiplex", "detection", "microsphere"], "article_id"=>692542, "categories"=>["Medicine", "Biological Sciences"], "users"=>["Ratthaphol Charlermroj", "Orawan Himananto", "Channarong Seepiban", "Mallika Kumpoosiri", "Nuchnard Warin", "Michalina Oplatowska", "Oraprapai Gajanandana", "Irene R. Grant", "Nitsara Karoonuthaisiri", "Christopher T. Elliott"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0062344.g002", "stats"=>{"downloads"=>0, "page_views"=>12, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Selection_of_antibody_pairs_for_the_multiplex_detection_using_a_microsphere_immunoassay_/692542", "title"=>"Selection of antibody pairs for the multiplex detection using a microsphere immunoassay.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-04-26 00:42:22"}
  • {"files"=>["https://ndownloader.figshare.com/files/1044823"], "description"=>"<div><p>Plant pathogens are a serious problem for seed export, plant disease control and plant quarantine. Rapid and accurate screening tests are urgently required to protect and prevent plant diseases spreading worldwide. A novel multiplex detection method was developed based on microsphere immunoassays to simultaneously detect four important plant pathogens: a fruit blotch bacterium <i>Acidovorax avenae</i> subsp. <i>citrulli</i> (Aac), chilli vein-banding mottle virus (CVbMV, potyvirus), watermelon silver mottle virus (WSMoV, tospovirus serogroup IV) and melon yellow spot virus (MYSV, tospovirus). An antibody for each plant pathogen was linked on a fluorescence-coded magnetic microsphere set which was used to capture corresponding pathogen. The presence of pathogens was detected by R-phycoerythrin (RPE)-labeled antibodies specific to the pathogens. The assay conditions were optimized by identifying appropriate antibody pairs, blocking buffer, concentration of RPE-labeled antibodies and assay time. Once conditions were optimized, the assay was able to detect all four plant pathogens precisely and accurately with substantially higher sensitivity than enzyme-linked immunosorbent assay (ELISA) when spiked in buffer and in healthy watermelon leaf extract. The assay time of the microsphere immunoassay (1 hour) was much shorter than that of ELISA (4 hours). This system was also shown to be capable of detecting the pathogens in naturally infected plant samples and is a major advancement in plant pathogen detection.</p></div>", "links"=>[], "tags"=>["Agricultural biotechnology", "crops", "Crop diseases", "Pest control", "Biochemistry", "immunochemistry", "biotechnology", "immunology", "Immunologic techniques", "immunoassays", "Plant science", "Plant pathology", "Plant pathogens", "detection", "pathogens", "microsphere", "immunoassay"], "article_id"=>692551, "categories"=>["Medicine", "Biological Sciences"], "users"=>["Ratthaphol Charlermroj", "Orawan Himananto", "Channarong Seepiban", "Mallika Kumpoosiri", "Nuchnard Warin", "Michalina Oplatowska", "Oraprapai Gajanandana", "Irene R. Grant", "Nitsara Karoonuthaisiri", "Christopher T. Elliott"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0062344", "stats"=>{"downloads"=>5, "page_views"=>56, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Multiplex_Detection_of_Plant_Pathogens_Using_a_Microsphere_Immunoassay_Technology_/692551", "title"=>"Multiplex Detection of Plant Pathogens Using a Microsphere Immunoassay Technology", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-04-26 00:42:31"}
  • {"files"=>["https://ndownloader.figshare.com/files/1044821"], "description"=>"<p><i>Acidovorax avenae</i> subsp. <i>citrulli</i> (Aac or A), potyvirus (P), watermelon silver mottle virus (WSMoV or W), capsicum chlorosis virus (CaCV or C) and melon yellow spot virus detection (MYSV or M) were used for finding the proper antibody pairs. Note: RPE is R-Phycoerythrin.</p>", "links"=>[], "tags"=>["Agricultural biotechnology", "crops", "Crop diseases", "Pest control", "Biochemistry", "immunochemistry", "biotechnology", "immunology", "Immunologic techniques", "immunoassays", "Plant science", "Plant pathology", "Plant pathogens", "antibody", "pairs", "multiplex"], "article_id"=>692550, "categories"=>["Medicine", "Biological Sciences"], "users"=>["Ratthaphol Charlermroj", "Orawan Himananto", "Channarong Seepiban", "Mallika Kumpoosiri", "Nuchnard Warin", "Michalina Oplatowska", "Oraprapai Gajanandana", "Irene R. Grant", "Nitsara Karoonuthaisiri", "Christopher T. Elliott"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0062344.t002", "stats"=>{"downloads"=>0, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Selection_of_antibody_pairs_for_the_multiplex_detection_/692550", "title"=>"Selection of antibody pairs for the multiplex detection.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-04-26 00:42:30"}
  • {"files"=>["https://ndownloader.figshare.com/files/1044818"], "description"=>"<p>The different concentrations of <i>Acidovorax avenae</i> subsp. <i>citrulli</i> (Aac) (A), recombinant coat protein (CP) of chilli vein-banding mottle virus (CVbMV) (B), recombinant nucleocapsid protein (NP) of watermelon silver mottle virus (WSMoV) (C) and melon yellow spot virus (MYSV) (D) were detected in the microsphere immunoassay using four incubation times: 15 min (circle), 30 min (square), 45 min (triangle) and 60 min (diamond). Y-axis is a median fluorescent intensity (MFI). Each data point was plotted as a mean of duplicates ± standard deviation. (E) Comparison of sensitivity of detection between microsphere immunoassay (four different incubation times) and sandwich ELISA (60 min incubation only) with the same sets of antibodies.</p>", "links"=>[], "tags"=>["Agricultural biotechnology", "crops", "Crop diseases", "Pest control", "Biochemistry", "immunochemistry", "biotechnology", "immunology", "Immunologic techniques", "immunoassays", "Plant science", "Plant pathology", "Plant pathogens", "assay"], "article_id"=>692547, "categories"=>["Medicine", "Biological Sciences"], "users"=>["Ratthaphol Charlermroj", "Orawan Himananto", "Channarong Seepiban", "Mallika Kumpoosiri", "Nuchnard Warin", "Michalina Oplatowska", "Oraprapai Gajanandana", "Irene R. Grant", "Nitsara Karoonuthaisiri", "Christopher T. Elliott"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0062344.g005", "stats"=>{"downloads"=>0, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effects_of_assay_time_on_sensitivity_of_detection_/692547", "title"=>"Effects of assay time on sensitivity of detection.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-04-26 00:42:27"}
  • {"files"=>["https://ndownloader.figshare.com/files/1044805"], "description"=>"<p>(A) The specific antibody-coated microspheres were mixed samples and incubated. (B) The unbound antigens were washed and removed by using magnetic separator. (C) The cocktail of RPE-labeled antibodies was added and incubated. (D) The unbound RPE-labeled antibodies were washed and removed by using magnetic separator before signals acquired by Luminex machine.</p>", "links"=>[], "tags"=>["Agricultural biotechnology", "crops", "Crop diseases", "Pest control", "Biochemistry", "immunochemistry", "biotechnology", "immunology", "Immunologic techniques", "immunoassays", "Plant science", "Plant pathology", "Plant pathogens", "microsphere"], "article_id"=>692540, "categories"=>["Medicine", "Biological Sciences"], "users"=>["Ratthaphol Charlermroj", "Orawan Himananto", "Channarong Seepiban", "Mallika Kumpoosiri", "Nuchnard Warin", "Michalina Oplatowska", "Oraprapai Gajanandana", "Irene R. Grant", "Nitsara Karoonuthaisiri", "Christopher T. Elliott"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0062344.g001", "stats"=>{"downloads"=>0, "page_views"=>13, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Scheme_of_magnetic_microsphere_immunoassay_/692540", "title"=>"Scheme of magnetic microsphere immunoassay.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-04-26 00:42:20"}
  • {"files"=>["https://ndownloader.figshare.com/files/1044820"], "description"=>"<p>Antibodies used in the study.</p>", "links"=>[], "tags"=>["Agricultural biotechnology", "crops", "Crop diseases", "Pest control", "Biochemistry", "immunochemistry", "biotechnology", "immunology", "Immunologic techniques", "immunoassays", "Plant science", "Plant pathology", "Plant pathogens"], "article_id"=>692549, "categories"=>["Medicine", "Biological Sciences"], "users"=>["Ratthaphol Charlermroj", "Orawan Himananto", "Channarong Seepiban", "Mallika Kumpoosiri", "Nuchnard Warin", "Michalina Oplatowska", "Oraprapai Gajanandana", "Irene R. Grant", "Nitsara Karoonuthaisiri", "Christopher T. Elliott"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0062344.t001", "stats"=>{"downloads"=>0, "page_views"=>10, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Antibodies_used_in_the_study_/692549", "title"=>"Antibodies used in the study.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-04-26 00:42:29"}
  • {"files"=>["https://ndownloader.figshare.com/files/1044814"], "description"=>"<p><i>Acidovorax avenae</i> subsp. <i>citrulli</i> (Aac) (10<sup>8</sup> CFU mL<b><sup>−</sup></b><sup>1</sup>), chilli vein-banding mottle virus (CVbMV) (0.2 µg mL<b><sup>−</sup></b><sup>1</sup>), watermelon silver mottle virus (WSMoV) (5 µg mL<b><sup>−</sup></b><sup>1</sup>), melon yellow spot virus (MYSV) (10 µg mL<b><sup>−</sup></b><sup>1</sup>) and mixed pathogens (10<sup>8</sup> CFU mL<b><sup>−</sup></b><sup>1</sup>) Aac, 0.2 µg mL<b><sup>−</sup></b><sup>1</sup> CVbMV, 5 µg mL<b><sup>−</sup></b><sup>1</sup> WSMoV and 10 µg mL<b><sup>−</sup></b><sup>1</sup> MYSV) in (A) 1% casein in PBST and (B) artificially spiked healthy watermelon leaf extract were tested using immuno microsphere. Antibody (MPC, 1B4, 2D6 and 5E7) coated microspheres were used to detect Aac, CVbMV, WSMoV and MYSV, respectively. Normalized signal (Y-axis) is a ratio of signal obtained from pathogen detection in the samples to the signal obtained when no pathogen was present. Each dataset was plotted as a mean of duplicates ± standard deviation.</p>", "links"=>[], "tags"=>["Agricultural biotechnology", "crops", "Crop diseases", "Pest control", "Biochemistry", "immunochemistry", "biotechnology", "immunology", "Immunologic techniques", "immunoassays", "Plant science", "Plant pathology", "Plant pathogens", "detection"], "article_id"=>692545, "categories"=>["Medicine", "Biological Sciences"], "users"=>["Ratthaphol Charlermroj", "Orawan Himananto", "Channarong Seepiban", "Mallika Kumpoosiri", "Nuchnard Warin", "Michalina Oplatowska", "Oraprapai Gajanandana", "Irene R. Grant", "Nitsara Karoonuthaisiri", "Christopher T. Elliott"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0062344.g004", "stats"=>{"downloads"=>0, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Multiplex_detection_of_four_plant_pathogens_/692545", "title"=>"Multiplex detection of four plant pathogens.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-04-26 00:42:25"}

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Relative Metric

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