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传统与微波组织处理技术对DNA完整性的影响:一项比较分子分析

Effect of Conventional and Microwave Tissue Processing Technique on DNA Integrity: A Comparative Molecular Analysis.

作者信息

Dwivedi Dhara, Kasetty Sowmya, Tijare Manisha S, Kallianpur Shreenivas, Prabhakar Nitin, Ragavendra Raju T, Desai Ami

机构信息

Department of Oral Pathology Dentistry Unit, Ayder Referral Hospital-College of Health Sciences Mekelle University, Ethiopia.

Oral pathology Division, Oral Basic and Clinical Sciences, College of Dentistry, Qassim Private College, Buraidah, KSA.

出版信息

Ethiop J Health Sci. 2018 Sep;28(5):615-624. doi: 10.4314/ejhs.v28i5.13.

Abstract

BACKGROUND

Methods of diagnostic molecular biology are routinely applied on formalin-fixed, paraffin-embedded tissues processed via conventional method. Recently, there has been a growing interest to use microwave technology in histopathology laboratories to overcome the deficiencies of the conventional processing method. Thefore, this study was aimed to compare and analyze the quality and quantity of DNA obtained from tissues processed by conventional and microwave tissue processing techniques and to further ascertain the applicability of the latter for PCR (polymerase chain reaction based research).

METHODS

Thirty fresh tissues of oral squamous cell carcinoma (OSCC) were included, and each sample was cut into two equivalent halves. One tissue half was processed by conventional manual method whereas the other half was processed using a domestic microwave oven. DNA was obtained from all the tissues which were then subjected to Polymerase chain reaction (PCR) to evaluate GAPDH (Glyceraldehyde-3-phosphate dehydrogenase) gene expression.

RESULTS

The results revealed better DNA yield from microwave processed tissue while the quality of the DNA was alike from both the techniques.

CONCLUSION

On the basis of the results obtained, it can be concluded that DNA produced by microwave processed tissues was similar to that obtained by conventional processing technique in terms of quantity and quality. Thus, microwave processed tissue samples can be successfully used for further molecular studies and researches.

摘要

背景

诊断分子生物学方法通常应用于通过传统方法处理的福尔马林固定、石蜡包埋组织。最近,在组织病理学实验室中使用微波技术以克服传统处理方法的不足的兴趣日益浓厚。因此,本研究旨在比较和分析通过传统和微波组织处理技术处理的组织所获得的DNA的质量和数量,并进一步确定后者在基于聚合酶链反应(PCR)的研究中的适用性。

方法

纳入30例口腔鳞状细胞癌(OSCC)新鲜组织,每个样本切成两个相等的部分。其中一半组织采用传统手工方法处理,另一半使用家用微波炉处理。从所有组织中提取DNA,然后进行聚合酶链反应(PCR)以评估甘油醛-3-磷酸脱氢酶(GAPDH)基因表达。

结果

结果显示,微波处理组织的DNA产量更高,而两种技术所获得的DNA质量相似。

结论

根据所获得的结果,可以得出结论,微波处理组织产生的DNA在数量和质量方面与传统处理技术所获得的DNA相似。因此,微波处理的组织样本可成功用于进一步的分子研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7242/6308768/509e41e71432/EJHS2805-0615Fig1.jpg

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