核酸检测试剂冻干技术及其评价方法研究进展

doi:10.19586/j.2095-2341.2025.0065

生物技术进展 ›› 2025, Vol. 15 ›› Issue (5): 804-811.DOI: 10.19586/j.2095-2341.2025.0065

核酸检测试剂冻干技术及其评价方法研究进展

刘莹莹¹(), 李若薇¹^,², 王艺霏¹

^1.中国质量检验检测科学研究院卫生检验与检疫研究所，北京 100176
^2.河北北方学院医学检验学院，河北张家口 075000

收稿日期:2025-05-26 接受日期:2025-07-30 出版日期:2025-09-25 发布日期:2025-11-11
作者简介:刘莹莹E-mail: liuyybd@126.com
基金资助:
中国质量检验检测科学研究院基本科研业务费项目(2024JK047)

Research Progress on Freeze-drying Technology and Evaluation Methods of Nucleic Acid Detection Reagents

Yingying LIU¹(), Ruowei LI¹^,², Yifei WANG¹

^1.Institute of Health Inspection and Quarantine，Chinese Academy of Quality and Inspection & Testing，Beijing 100176，China
^2.School of Medical Laboratory Medicine，Hebei North University，Hebei Zhangjiakou 075000，China

Received:2025-05-26 Accepted:2025-07-30 Online:2025-09-25 Published:2025-11-11

摘要/Abstract

摘要：

核酸检测在疾病诊断、治疗指导、及时防控以及科学研究与公共卫生监测等多个领域扮演着至关重要的角色。然而，传统的液体试剂在储存和运输过程中面临诸多挑战，如稳定性差、灵敏度下降等问题。随着冻干技术的进步，其在核酸检测试剂中的应用取得了显著成效。综述了冻干技术在核酸检测试剂中的应用进展，重点探讨了冻干保护剂的选择与浓度优化、冻干条件对试剂性能的影响、冻干后试剂形貌调控以及冻干试剂检测性能与稳定性的评价方法，旨在为后续相关研究提供科学指导，推动冻干技术在临床诊断、疾病预防和生物科学研究中的应用。

关键词: 核酸检测, 冻干技术, 冻干保护剂, 评价方法

Abstract:

Nucleic acid testing plays a crucial role in disease diagnosis， treatment guidance， timely prevention and control， as well as scientific research and public health monitoring. However， traditional liquid reagents face numerous challenges during storage and transportation， such as poor stability and reduced sensitivity. With advancements in freeze-drying technology， its application in nucleic acid testing reagents has achieved significant results. The review summarized the progress of freeze-drying technology in nucleic acid testing reagents， focusing on the selection and concentration optimization of freeze-drying protectants， the impact of freeze-drying conditions on reagent performance， the regulation of reagent morphology after freeze-drying， and evaluation methods for the detection performance and stability of freeze-dried reagents. This paper aims to provide scientific guidance for subsequent related research and promote the application of freeze-drying technology in clinical diagnosis， disease prevention， and biological science research.

Key words: nucleic acid testing, lyophilization technology, lyophilized protective agent, evaluation methods

中图分类号:

Q945

刘莹莹, 李若薇, 王艺霏. 核酸检测试剂冻干技术及其评价方法研究进展[J]. 生物技术进展, 2025, 15(5): 804-811.

Yingying LIU, Ruowei LI, Yifei WANG. Research Progress on Freeze-drying Technology and Evaluation Methods of Nucleic Acid Detection Reagents[J]. Current Biotechnology, 2025, 15(5): 804-811.

图/表 2

参考文献 61

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分类	名称
糖类	蔗糖、海藻糖、葡萄糖、乳糖、麦芽糖、果聚糖
醇类	甘油、甘露醇、山梨醇、肌醇、木糖醇
氨基酸类	甘氨酸、精氨酸、组氨酸、色氨酸、丙氨酸、丝氨酸
聚合物类	聚乙烯吡咯烷酮、聚乙二醇、牛血清白蛋白、右旋糖酐
表面活性剂	吐温80、曲拉通X-100、十二烷基硫酸钠
盐类	磷酸盐、枸橼酸盐、柠檬酸盐、醋酸盐、氯化钠

分类	名称
糖类	蔗糖、海藻糖、葡萄糖、乳糖、麦芽糖、果聚糖
醇类	甘油、甘露醇、山梨醇、肌醇、木糖醇
氨基酸类	甘氨酸、精氨酸、组氨酸、色氨酸、丙氨酸、丝氨酸
聚合物类	聚乙烯吡咯烷酮、聚乙二醇、牛血清白蛋白、右旋糖酐
表面活性剂	吐温80、曲拉通X-100、十二烷基硫酸钠
盐类	磷酸盐、枸橼酸盐、柠檬酸盐、醋酸盐、氯化钠

评价方法	评价内容	优点	缺点
物理评价	观察冻干试剂的颜色、形态等物理性质，确保其未发生明显变化；检测冻干试剂的溶解速度，评估其复溶性能	直观、简单，能够快速筛选出外观异常或溶解性能不佳的试剂	无法全面反映试剂的生物活性和化学稳定性，需要结合其他方法进行综合评估
检测性能评价	通过电泳法，比较冻干前后检测试剂扩增产物目标条带明亮程度；通过荧光定量PCR技术，比较冻干前后检测试剂扩增曲线变化，C_t 值变化进行评估	能够直接反映试剂对样本的检测灵敏度，是评估试剂质量的重要指标	需要专业的实验条件和操作技能，且实验周期较长，成本较高
稳定性评价	通过多次重复实验，计算冻干试剂检测结果的准确性、精密度、特异性等指标；评估冻干试剂在存在干扰物的情况下的检测性能，确保其在复杂样本中的准确性	能够提供客观、量化的评估结果，有助于比较不同试剂之间的性能差异	周期较长且需要大量的实验数据和统计分析技术，对实验设计和数据处理能力要求较高

评价方法	评价内容	优点	缺点
物理评价	观察冻干试剂的颜色、形态等物理性质，确保其未发生明显变化；检测冻干试剂的溶解速度，评估其复溶性能	直观、简单，能够快速筛选出外观异常或溶解性能不佳的试剂	无法全面反映试剂的生物活性和化学稳定性，需要结合其他方法进行综合评估
检测性能评价	通过电泳法，比较冻干前后检测试剂扩增产物目标条带明亮程度；通过荧光定量PCR技术，比较冻干前后检测试剂扩增曲线变化，C_t 值变化进行评估	能够直接反映试剂对样本的检测灵敏度，是评估试剂质量的重要指标	需要专业的实验条件和操作技能，且实验周期较长，成本较高
稳定性评价	通过多次重复实验，计算冻干试剂检测结果的准确性、精密度、特异性等指标；评估冻干试剂在存在干扰物的情况下的检测性能，确保其在复杂样本中的准确性	能够提供客观、量化的评估结果，有助于比较不同试剂之间的性能差异	周期较长且需要大量的实验数据和统计分析技术，对实验设计和数据处理能力要求较高

核酸检测试剂冻干技术及其评价方法研究进展

Research Progress on Freeze-drying Technology and Evaluation Methods of Nucleic Acid Detection Reagents

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