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Information Resource
Record information and status
Record ID
115229
Status
Published
Date of creation
2019-10-21 16:56 UTC (austein.mcloughlin@cbd.int)
Date of publication
2019-10-21 16:57 UTC (austein.mcloughlin@cbd.int)

General Information
Title
A novel emulsion PCR method coupled with fluorescence spectrophotometry for simultaneous qualitative, quantitative and high-throughput detection of multiple DNA targets
Author
Yanan Du, Xiao Zhao, Binan Zhao, Yan Xu, Wei Shi, Fangfang Ren, Yangyang Wu, Ruili Hu, Xiaorui Fan, Qi Zhang, Xiaoxia Zhang, Wanjing Zhang, Weijing Wu, Bin Shi, Huanzhen Zhao, Kai Zhao
Language(s)
  • English
Publication date
2019-01-19
Subject
Summary, abstract or table of contents

A novel emulsion PCR method combined with fluorescence spectrophotometry (EPFS) for simultaneous qualitative, quantitative and high-throughput detection of multiple DNA targets was tested and validated. In a single reaction set, each pair of primers was labeled with a specific fluorophore. Through emulsion PCR, a target DNA was amplified in droplets that functioned as microreactors. After product purification, different fluorescent-labeled DNA products were qualitatively analyzed by the fluorescent intensity determination. The sensitivity and specificity of the system was examined using four kinds of modified maize. The qualitative results revealed high specificity and sensitivity of 0.5% (w/w). In addition, the quantitative results revealed that the absolute limit of detection was 103 copies, showing good repeatability. Moreover, the reproducibility assays were further performed using four foodborne pathogenic bacteria to further evaluate the applicability of the system. Consequently, the same qualitative, quantitative and high-throughput results were confirmed with the four GM maize. Overall, the new EPFS system is an analytical technology that  could enable simultaneous qualitative, quantitative and high-throughput analysis of multiple genes.
Thematic areas
Information on Organisms or LMOs
LMO(s) identification
SYN-EV176-9 - NaturGard KnockOut™ maize
Resistance to antibiotics - Ampicillin Resistance to diseases and pests - Insects - Lepidoptera (butterflies and moths) Resistance to herbicides - Glufosinate
Show detection method(s)
MON-ØØØ21-9 - Roundup Ready™ maize
Resistance to herbicides - Glyphosate
Show detection method(s)
MON-ØØ6Ø3-6 - Roundup Ready™ maize
Resistance to herbicides - Glyphosate
Show detection method(s)
DAS-Ø15Ø7-1 - Herculex™ I maize
Resistance to diseases and pests - Insects - Lepidoptera (butterflies and moths) Resistance to herbicides - Glufosinate
Show detection method(s)
Additional Information
Type of resource
  • Article (journal / magazine / newspaper)
Identifier
DOI:10.1038/s41598-018-36981-1
Publisher and its location
Scientific Reports
Format
PDF file (9 pages) - 1223.5kB
Source
Online article from journal
Keywords and any other relevant information
Detection, Identification, maize, emulsion PCR, quantitative