Detection on genetically modified soybean by helicase-dependent isothermal DNA amplification assay

被引：0
|
作者：
Zhang J. [1 ]
Wang Z. [1 ]
Zhang C. [1 ]
Ma C. [1 ]
Zhou W. [1 ,2 ]
Zhang Y. [1 ]
机构：
[1] Hebei Food Safety Key Laboratory, Hebei Food Inspection and Research Institute, Shijiazhuang
[2] College of Food Science and Technology, Agricultural University of Hebei, Baoding
来源：
Shipin Kexue/Food Science | 2016年 / 37卷 / 04期
关键词：
Detection; Genetically modified soybean; Helicase-dependent isothermal DNA amplification (HDA);
D O I：
10.7506/spkx1002-6630-201604029
中图分类号：
学科分类号：
摘要：
Objective: To establish a new rapid method to detect genetically modified soybean based on helicase-dependent isothermal DNA amplification (HDA). Methods: With four highly specific sets of primers synthesized to target genes including Lectin in soybean and CaMV35S, NOS and CP4-EPSPS foreign genes, PCR reaction system was established. The specificity and sensitivity of the PCR method were tested based on HDA and homology analysis was performed. Results: The helicase-dependent isothermal DNA amplification method for the rapid detection of genetically modified soybean has been established. The detection limits of CaMV35S, NOS and CP4-EPSPS genes in genetically modified soybeans by HDA were all 0.2%. Conclusion: HDA is a rapid, user-friendly, specific and sensitive method for the detection of genetically modified soybean, and is very suitable for use in grassroots laboratories with a broad prospect. © 2016, China Food Publishing Company. All right reserved.
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页码：164 / 168
页数：4
相关论文
共 12 条
  • [1] Lerat S., England L.S., Vincent M.L., Et al., Real-time polymerase chain reaction quantification of the transgenes for roundup ready corn and roundup ready soybean in soil samples, Journal of Agricultural and Food Chemistry, 53, 5, pp. 1337-1342, (2005)
  • [2] Cao Y., Kim H.J., Li Y., Et al., Helicase-dependent amplification of nucleic acids, Current Protocols in Molecular Biology, 104, pp. 1511-1512, (2013)
  • [3] Chen X., Wu X., Gan M., Et al., Rapid detection of Staphylococcus aureus in dairy and meat foods by combination of capture with silicacoated magnetic nanoparticles and thermophilic helicase-dependent isothermal amplification, Journal of Dairy Science, 98, 3, pp. 1563-1570, (2015)
  • [4] Barreda-Garcia S., Gonzalez-Alvarez M.J., Delos-Santos-Alvarez N., Et al., Attomolar quantitation of Mycobacterium tuberculosis by asymmetric helicase-dependent isothermal DNA-amplification and electrochemical detection, Biosens Bioelectron, 68, pp. 122-128, (2014)
  • [5] Li Y., Kumar N., Gopalakrishnan A., Et al., Detection and species identification of malaria parasites by isothermal tHDA amplification directly from human blood without sample preparation, Journal of Molecular Diagnostics, 15, 5, pp. 634-641, (2013)
  • [6] Huang S., Do J., Mahalanabis M., Et al., Low cost extraction and isothermal amplification of DNA for infectious diarrhea diagnosis, PLoS One, 8, 3, (2013)
  • [7] Eckert C., Holscher E., Petit A., Et al., Molecular test based on isothermal helicase-dependent amplification for detection of the Clostridium difficile toxin A gene, Journal of Clinical Microbiology, 52, 7, pp. 2386-2389, (2014)
  • [8] Moura-Melo S., Miranda-Castro R., De-Los-Santosalvarez N., Et al., Targeting helicase-dependent amplification products with an electrochemical genosensor for reliable and sensitive screening of genetically-modified organisms, Analytical Chemistry, 87, 16, pp. 8547-8554, (2015)
  • [9] Thongsri Y., Wonglakorn L., Chaiprasert A., Et al., Evaluation for the clinical diagnosis of Pythium insidiosum using a single-tube nested PCR, Mycopathologia, 176, 5-6, pp. 369-376, (2013)
  • [10] Barbieri D., Venturoli S., Rosl F., Et al., Detection of high-risk human papillomavirus type 16 and 18 using isothermal helicase-dependent amplification, Diagnostic Microbiology and Infectious Disease, 79, 2, pp. 178-182, (2014)
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