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Rapid identification of floral origin in honey by single strand DNA-binding protein-enhanced denaturation bubble-mediated strand exchange amplification
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Qian WU1, 2, Jin-Tian YANG2, Jian ZHANG3, Feng-Xia SUN3, De-Xin ZENG4, Wei CHEN2, *, Xiao-Feng YU4, *
Journal of Food Safety & Quality | 2026, 17(10) : 130 - 139
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Journal of Food Safety & Quality | 2026, 17(10): 130-139
• Special Topic: Food Authenticity and Traceability •
Rapid identification of floral origin in honey by single strand DNA-binding protein-enhanced denaturation bubble-mediated strand exchange amplification
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Qian WU1, 2, Jin-Tian YANG2, Jian ZHANG3, Feng-Xia SUN3, De-Xin ZENG4, Wei CHEN2, *, Xiao-Feng YU4, *
Affiliations
  • 1.College of Food and Bioengineering, Bengbu University, Bengbu 233000, China
  • 2.School of Food Science &Biological Engineering, Hefei University of Technology, Hefei 230000, China
  • 3.School of Food Science & Technology,Shihezi University, Shihezi 832003, China
  • 4.Technical Center of Hefei Custom, Hefei 230000, China
doi: 10.19812/j.cnki.jfsq11-5956/ts.20251205007
Outline
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Objective

To establish a rapid identification method for the authenticity of honey botanical origin,based on an isothermal amplification approach for honey pollen gene detection, combined with a time-resolved fluorescent microsphere lateral flow strip (TR-LFS).

Methods

A rapid DNA extraction method for honey pollen was constructed to obtain the effective maturase K (matK) target gene template of Robinia pseudoacacia within 20 min.Denaturation bubble-mediated strand exchange amplification (SEA) was employed to achieve exponential amplification of the target sequence under a constant temperature of approximately 65 °C. To enhance amplification efficiency, single strand DNA-binding protein (SSB) was added to inhibit non-specific annealing of single strands,significantly improving primer hybridization efficiency and amplification specificity. When target pollen DNA was present in the honey, the SEA amplification generated double-stranded products labeled at both ends, which were subsequently detected visually via the TR-LFS based on fluorescence.

Results

The fluorescence signal intensity of the TR-LFS allowed for visual determination of honey botanical origin authenticity. Using Robinia pseudoacacia honey as a model, the naked-eye detection sensitivity of this method for the target content reached 1% (V:V), and no cross-reaction was observed with other botanical honey sources.

Conclusion

The detection method established in this study significantly enhances the amplification efficiency of SEA technology through the introduction of SSB.Combined with the high sensitivity and stable fluorescence output characteristics of TR-LFS, and eliminating the need for complex thermal cycling equipment, this method is suitable for on-site rapid detection, providing reliable technical support for authenticating honey floral sources.

honey authenticity  /  molecular amplification  /  single strand DNA-binding protein  /  lateral flow strip  /  on-site rapid detection
吴倩, 杨金田, 张建, 孙凤霞, 曾德新, 陈伟, 余晓峰. 单链DNA结合蛋白辅助增强型变性泡介导链交换扩增法快速鉴定蜂蜜花源成分. 食品安全质量检测学报, 2026 , 17 (10) : 130 -139 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20251205007
Qian WU, Jin-Tian YANG, Jian ZHANG, Feng-Xia SUN, De-Xin ZENG, Wei CHEN, Xiao-Feng YU. Rapid identification of floral origin in honey by single strand DNA-binding protein-enhanced denaturation bubble-mediated strand exchange amplification[J]. Journal of Food Safety & Quality, 2026 , 17 (10) : 130 -139 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20251205007
Appendix
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Year 2026 volume 17 Issue 10
7
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Article Info
doi: 10.19812/j.cnki.jfsq11-5956/ts.20251205007
  • Receive Date：2025-12-05
  • Online Date：2026-11-05
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History
  • Received：2025-12-05
Funding
Affiliations
    1.College of Food and Bioengineering, Bengbu University, Bengbu 233000, China
    2.School of Food Science &Biological Engineering, Hefei University of Technology, Hefei 230000, China
    3.School of Food Science & Technology,Shihezi University, Shihezi 832003, China
    4.Technical Center of Hefei Custom, Hefei 230000, China
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表12种不同金属材料的力学参数
科
Family		 属数
Number of
genus		 种数
Number of
species		 占总种数比例
Percentage of
total species (%)		 属
Genus		 种数
Number of
species		 占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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