Dataset Information

Detection of GM Canola MS11, DP-073496-4, and MON88302 events using multiplex PCR coupled with capillary electrophoresis.

ABSTRACT: As of 2020, 11 GM canola events have been authorized as food for humans in Korea. However, there are no simultaneous multiplex detection methods for 3 GM canola events (DP-073496-4, MON88302, and MS11). Thus, we established the multiplex polymerase chain reaction (PCR) method coupled with capillary electrophoresis to detect 3 GM canola events. To verify the specificity of event-specific primers, various GM crops of 3 GM soybean events, 6 GM maize events, 2 GM cotton events and 11 GM canola events were prepared. The limit of detection of the developed multiplex PCR was approximately 0.0125% for 3 GM canola events. Certified GM canola and stacked events were analyzed to validate the developed multiplex PCR. This study focuses on establishing multiplex PCR coupled with capillary electrophoresis for newly approved GM canola events and contributes to efficient monitoring GM canola samples in Korea.

SUBMITTER: Lee DG

PROVIDER: S-EPMC8050189 | biostudies-literature | 2021 Apr

REPOSITORIES: biostudies-literature

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Publications

Detection of GM Canola MS11, DP-073496-4, and MON88302 events using multiplex PCR coupled with capillary electrophoresis.

Lee Do-Geun DG Park Ji-Eun JE Kim Mi-Ju MJ Kim Hyun-Joong HJ Kim Hae-Yeong HY

Food science and biotechnology 20210313 4

As of 2020, 11 GM canola events have been authorized as food for humans in Korea. However, there are no simultaneous multiplex detection methods for 3 GM canola events (DP-073496-4, MON88302, and MS11). Thus, we established the multiplex polymerase chain reaction (PCR) method coupled with capillary electrophoresis to detect 3 GM canola events. To verify the specificity of event-specific primers, various GM crops of 3 GM soybean events, 6 GM maize events, 2 GM cotton events and 11 GM canola event ...[more]

PMID: 33936848

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Project description:Cost-effective molecular diagnostic techniques for bacterial pneumonia are limited. We designed primers for 13 bacteria, performed multiplex nucleic acid detection through fragment analysis to obtain pathogen identification results, and established a multiplex PCR-capillary electrophoresis (MPCE) method, which can simultaneously detect 13 pathogens associated with bacterial pneumonia. The sensitivity, specificity, and reproducibility of the MPCE assay were tested, and 420 clinical samples were used to assess the clinical detection ability of MPCE, with the culture method used as a reference. Samples with inconsistent results detected by the two methods were sent for Sanger sequencing. The minimum detection limit of MPCE for 13 bacteria was 6.0 × 103 cfu/mL~2.0 × 106 cfu/mL. No cross-reactivity was observed with other pathogens. The percentage of agreement for reproducibility analysis reached 100%. For the 420 sputum samples, when the culture method was used as the reference, the sensitivity of MPCE to 13 bacteria ranged from 80% to 100%. The specificity for 13 bacteria ranged from 67.1% to 100%. The percentage of agreement between the MPCE and the culture method ranged from 69.7% to 100%. There was no statistically significant difference (P > 0.05) in the detection of Escherichia coli, Enterobacter cloacae complex, Staphylococcus aureus, methicillin-resistant S. aureus, Streptococcus pyogenes, Moraxella catarrhalis, or Legionella pneumophila between the MPCE and the culture method. Clinical samples with negative cultures but positive MPCE results were validated by Sanger sequencing, and the results were consistent with those of MPCE. The MPCE method has high sensitivity and specificity for bacterial pneumonia, enabling the simultaneous and rapid detection of multiple pathogens. It is cost-effective and has potential for clinical application.ImportanceThis study successfully established a multiplex PCR-capillary electrophoresis detection system that can simultaneously detect 13 pathogens through a single detection method, significantly improving clinical efficiency. It is cost-effective and has potential for clinical application.

Dataset Information

Detection of GM Canola MS11, DP-073496-4, and MON88302 events using multiplex PCR coupled with capillary electrophoresis.

Publications

Detection of GM Canola MS11, DP-073496-4, and MON88302 events using multiplex PCR coupled with capillary electrophoresis.

Similar Datasets

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