组织保护剂对病毒核酸与豚鼠皮肤组织样本RNA的保存效果探究

doi:10.19586/j.2095-2341.2023.0123

生物技术进展 ›› 2024, Vol. 14 ›› Issue (1): 60-65.DOI: 10.19586/j.2095-2341.2023.0123

• 生物制品研发与技术专题 • 上一篇下一篇

组织保护剂对病毒核酸与豚鼠皮肤组织样本RNA的保存效果探究

李华¹(), 徐兰举¹, 黄学亮², 王亚如³, 刁立琴³(), 于月欣¹()

^1.河北纳科生物科技有限公司，石家庄 050035
^2.河北以岭医院，石家庄 050090
^3.河北省药品医疗器械检验研究院，石家庄 050227

收稿日期:2023-10-13 接受日期:2023-12-21 出版日期:2024-01-25 发布日期:2024-02-05
通讯作者: 刁立琴,于月欣
作者简介:李华E-mail: 15909835232@163.com
基金资助:
河北省引进国外智力项目

Preservation Effect of Tissue Protectant on Viral Nucleic Acid and RNA of Guinea Pig Skin Tissue Samples

Hua LI¹(), Lanju XU¹, Xueliang HUANG², Yaru WANG³, Liqin DIAO³(), Yuexin YU¹()

^1.Hebei NACOL Biotechnology Co. ，Ltd. ，Shijiazhuang 050035，China
^2.Hebei Yiling Hospital，Shijiazhuang 050090，China
^3.Hebei Institute for Drug and Medical Device Control，Shijiazhuang 050227，China

Received:2023-10-13 Accepted:2023-12-21 Online:2024-01-25 Published:2024-02-05
Contact: Liqin DIAO,Yuexin YU

摘要/Abstract

摘要：

病毒核酸和生物组织样本RNA容易受到外部环境的影响，极易发生降解，造成实验结果产生严重偏差。以病毒核酸与豚鼠皮肤组织样本RNA为研究对象，探究组织保护剂保存它们在不同温度放置不同时间后，对病毒载量以及豚鼠皮肤组织样本中RNA的保护效果。结果发现，在4 ℃保存30 d，25 ℃保存7 d或37 ℃保存24 h后，与初始病毒核酸量相比较，组织保护剂中保存的病毒载量未发生明显变化（P>0.05）。在4 ℃保存30 d，25 ℃保存7 d或37 ℃保存24 h后，与液氮储存组相比较，组织保护剂组与市售RNA later组均可以保持豚鼠皮肤组织中样本RNA的提取量和纯度，差异无统计学意义（P>0.05）。结果表明，组织保护剂可作为科研或临床组织样本的储存保存液，在4 ℃、25 ℃以及37 ℃条件下放置一定时间，不影响病毒核酸的病毒载量以及豚鼠皮肤组织样本RNA的提取量和纯度，使样本保存更简便高效。

关键词: 组织保护剂, 病毒载量, 组织样本RNA, 保护效果

Abstract:

Viral nucleic acid and RNA of biological tissue samples are easy to be affected by external environment and easily degrade， resulting in serious deviations in experimental results. Taking viral nucleic acid and guinea pig skin tissue sample RNA as research objects， the protective effect of tissue protectant on viral load and RNA in guinea pig skin tissue sample after preservation of them at different temperatures and different times was explored. The results showed that after storage at 4 ℃ for 30 days （d）， 25 ℃ for 7 d or 37 ℃ for 24 hours （h）， the viral load stored in tissue protectants did not change significantly compared with the initial viral nucleic acid dose （P>0.05）. After storage at 4 ℃ for 30 d， 25 ℃ for 7 d or 37 ℃ for 24 h， compared with liquid nitrogen storage group， both tissue protectant group and commercially available RNA later group could maintain the extraction amount and purity of sample RNA in guinea pig skin tissue， with no statistical significance （P>0.05）. The results showed that the tissue protectant could be used as a storage solution for scientific research or clinical tissue samples. It was placed at 4 ℃， 25 ℃ and 37 ℃ for a certain time， without affecting the viral load of viral nucleic acid and the amount and purity of RNA extraction from guinea pig skin tissue samples， which made the sample preservation more convenient and efficient.

Key words: sample preservation solution, viral load, tissue sample RNA, protective effect

中图分类号:

李华, 徐兰举, 黄学亮, 王亚如, 刁立琴, 于月欣. 组织保护剂对病毒核酸与豚鼠皮肤组织样本RNA的保存效果探究[J]. 生物技术进展, 2024, 14(1): 60-65.

Hua LI, Lanju XU, Xueliang HUANG, Yaru WANG, Liqin DIAO, Yuexin YU. Preservation Effect of Tissue Protectant on Viral Nucleic Acid and RNA of Guinea Pig Skin Tissue Samples[J]. Current Biotechnology, 2024, 14(1): 60-65.

图/表 6

表1 4 ℃放置不同时间后病毒核酸载量的比较

Table 1 Comparison of viral nucleic acid load at 4 ℃ for different time

组别	0 d	7 d	14 d	21 d	30 d
Ct值	35.46±0.06	35.50±0.09	35.40±0.16	35.49±0.05	35.45±0.05
t	—	1.65	1.57	1.72	0.57
P	—	0.11， P>0.05	0.13， P>0.05	0.09， P>0.05	0.57， P>0.05

表2 25 ℃放置不同时间后病毒核酸载量的比较

Table 2 Comparison of viral nucleic acid load at 25 ℃ for different time

组别	0 d	1 d	3 d	5 d	7 d
Ct值	35.46±0.06	35.46±0.05	35.45±0.04	35.45±0.06	35.46±0.15
t	—	0.00	0.62	0.53	0.00
P	—	1.00， P>0.05	0.54， P>0.05	0.60， P>0.05	1.00， P>0.05

表3 37 ℃放置不同时间对病毒核酸载量的比较

Table 3 Comparison of viral nucleic acid load at 37 ℃ for different time

组别	0 h	6 h	12 h	18 h	24 h
Ct值	35.46±0.06	35.46±0.08	35.50±0.09	35.48±0.10	35.48±0.08
t	—	0.00	1.65	0.77	0.89
P	—	1.00， P>0.05	0.11， P>0.05	0.45， P>0.05	0.38， P>0.05

图1 4 ℃放置30 d后不同处理对RNA保护的验证结果A：各组提取的RNA提取总量；B：各组提取总RNA的琼脂糖凝胶图谱，M—DNA maker；C~D：各组提取的RNA纯度

Fig. 1 Verification results of RNA protection by tissue protectant after 30 days at 4 ℃

图2 25 ℃放置7 d后不同处理对RNA保护的验证结果A：各组提取的RNA提取总量；B：各组提取总RNA的琼脂糖凝胶图谱，M—DNA maker；C~D：各组提取的RNA纯度

Fig. 2 Verification results of RNA protection by tissue protectant after 7 days at 25 ℃

图3 37 ℃放置24 h后不同处理对RNA保护的验证结果A：各组提取的RNA提取总量；B：各组提取总RNA的琼脂糖凝胶图谱，M—DNA maker；C~D：各组提取的RNA纯度

Fig. 3 Verification results of RNA protection by tissue protectant after 24 hours at 37 ℃

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组织保护剂对病毒核酸与豚鼠皮肤组织样本RNA的保存效果探究

Preservation Effect of Tissue Protectant on Viral Nucleic Acid and RNA of Guinea Pig Skin Tissue Samples

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