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Study on the extraction of yeast mRNA and the detection method of reverse transcription real-time fluorescence quantitative polymerase chain reaction
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Yun-Xia LI, Rui ZHANG, Fang HAN, Sheng-Long MA, Ying-Ying NIE, Li-Ping MA*
Journal of Food Safety & Quality | 2026, 17(9) : 314 - 323
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Journal of Food Safety & Quality | 2026, 17(9): 314-323
• Food Analysis and Detection •
Study on the extraction of yeast mRNA and the detection method of reverse transcription real-time fluorescence quantitative polymerase chain reaction
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Yun-Xia LI, Rui ZHANG, Fang HAN, Sheng-Long MA, Ying-Ying NIE, Li-Ping MA*
Affiliations
  • Institute of Sensing Technology, Gansu Academy of Sciences, Lanzhou 730000, China
doi: 10.19812/j.cnki.jfsq11-5956/ts.20260104005
Outline
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Objective

To explore an efficient method for extracting messenger RNA (mRNA) from yeast and establish a specific detection method by reverse transcription real-time fluorescence quantitative polymerase chain reaction (RT-qPCR).

Methods

A high-performance silica magnetic beads by self-developed were used as carrier, an efficient yeast mRNA extraction technique was developed by combining streptavidin magnetic beads (SM) conjugated with Oligo(dT) and Trizol lysis, followed by optimization of magnetic bead concentration and magnetic adsorption time. Primers based on the yeast chitin synthase 2 (CHS2) gene, established a yeast-specific RT-qPCR assay, and its specificity and stability were validated.

Results

Under optimized extraction conditions, the method enabled extraction yeast mRNA from various concentration, yielded 2.13 μg mRNA even from a low-concentration (2×103 cfu/mL). This yeast detection methodexhibited a wide linear range from 2×103 to 2×107 cfu/mL, with a linear equation of Y=–1.69X+33.412 (r2=0.9966), the method exhibited high specificity for the target yeast.

Conclusion

This study successfully establish a mRNA extraction technique by yeast-specific magnetic bead-based and a RT-qPCR detection method, exhibits good specificity and sensitivity, provide a new technology for the detection of yeast contamination in food and scientific research.

yeasts  /  mRNA extraction  /  reverse transcription real-time fluorescence quantitative polymerase chain reaction  /  chitin synthase 2
李云霞, 张锐, 韩芳, 马生龙, 聂莹莹, 马莉萍. 酵母菌mRNA提取技术及实时荧光定量逆转录聚合酶链式反应检测方法研究. 食品安全质量检测学报, 2026 , 17 (9) : 314 -323 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20260104005
Yun-Xia LI, Rui ZHANG, Fang HAN, Sheng-Long MA, Ying-Ying NIE, Li-Ping MA. Study on the extraction of yeast mRNA and the detection method of reverse transcription real-time fluorescence quantitative polymerase chain reaction[J]. Journal of Food Safety & Quality, 2026 , 17 (9) : 314 -323 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20260104005
Appendix
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Year 2026 volume 17 Issue 9
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Article Info
doi: 10.19812/j.cnki.jfsq11-5956/ts.20260104005
  • Receive Date：2026-01-04
  • Online Date：2026-11-05
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History
  • Received：2026-01-04
Funding
Affiliations
    Institute of Sensing Technology, Gansu Academy of Sciences, Lanzhou 730000, 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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