Hajira Khatoon, N S Sahana, T Suresh, Jyoti Tahasildar, S Renuga, Meghashyama Kulkarni
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引用次数: 0
Abstract
Background: Analysis of mRNA in archival tissues using RT-qPCR has the potential to become an important element in diagnosis and research. There is uncertainty whether mRNA extraction and analyses from archival tissues are possible or not. This study will look for the possibility of mRNA extraction, RT-qPCR analysis, and standardization of the protocol using formalin fixed paraffin-embedded blocks.
Objectives: 1. To determine the effect of 24-hour and 72-hour formalin-fixed paraffin-embedded tissue on the quantity and quality of mRNA. 2. To compare the quantity and quality of mRNA in fresh frozen tissues. 3. To compare the extracted mRNA expression using RT-qPCR in the above groups.
Methodology: Twelve tissue samples were collected from patients undergoing minor surgical procedures and grossed into 3 bits. Each bit was placed in 24 hours of formalin and 72 hours of formalin, and the last bit was freezed in RNAlater at -80°C (positive controls), respectively. Routine tissue processing and sectioning was done followed by wax removal for the formalin-fixed tissues, and mRNA extraction using TRIzol method was done for all three groups. Extracted mRNA was quantified using Nanodrop spectrophotometer and its quality checked on mRNA TapeStation. cDNA synthesis was done followed by RT-qPCR analysis.
Results: mRNA could be isolated with satisfactory quantity in all three groups. mRNA quality was significantly low for formalin-fixed tissues. But the RT-qPCR values of the two formalin groups were comparable to those obtained in fresh frozen tissues (P value = 0.00002).
Conclusion: mRNA can be extracted from archives of paraffin tissue blocks that can be utilized to carry out enormous studies using RT-qPCR.
期刊介绍：
The journal of Oral and Maxillofacial Pathology [ISSN:print-(0973-029X, online-1998-393X)] is a tri-annual journal published on behalf of “The Indian Association of Oral and Maxillofacial Pathologists” (IAOMP). The publication of JOMFP was started in the year 1993. The journal publishes papers on a wide spectrum of topics associated with the scope of Oral and Maxillofacial Pathology, also, ensuring scientific merit and quality. It is a comprehensive reading material for the professionals who want to upgrade their diagnostic skills in Oral Diseases; allows exposure to newer topics and methods of research in the Oral-facial Tissues and Pathology. New features allow an open minded thinking and approach to various pathologies. It also encourages authors to showcase quality work done by them and to compile relevant cases which are diagnostically challenging. The Journal takes pride in maintaining the quality of articles and photomicrographs.
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