ISSN : 2663-2187

AN OVERVIEW ON RT-PCR METHODS FOR ESTIMATION OF CIRCULATING miRNA BREAST CANCER BIOMARKERS

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Villuri Ruth Beulah and B. Sailaja
ยป doi: 10.48047/AFJBS.7.1.2025.858-867

Abstract

Breast cancer is the most common cause of cancer-related fatalities in women, early diagnosis of breast cancer helps in decreasing the mortality, improving the survival rate and reducing the cost of health care. Early detection of breast cancer improves the 5year survival rate up to 100%. The normal function of microRNAs (miRNAs) is cell proliferation, differentiation and apoptosis in healthy individuals. Circulating breast cancer miRNAs are present in body fluids like serum, plasma, saliva, and urine. Breast cancer miRNA biomarkers are oncogenic or tumor suppressors. They are involved in tumor initiation and advancement and their concentrations are markedly altered in patients with breast cancer. Thus enable them to be used as diagnostic tools in breast cancer. As the miRNA biomarkers are present in low concentration, RT-PCR is the suitable technique and is widely used for accurately estimating miRNAs with high reproducibility. A more stable and sensitive RT-PCR method involves the inclusion of a panel of biomarker rather than a single biomarker. Reported studies suggest that miR-1246, miR-24, miR-373 panel is a superior, validated oncogenic biomarker panel with 98% sensitivity and 97% accuracy in stage I and II breast cancer patients. This review aspires to give an overview on different sample processing techniques, miRNA extraction kits used and qPCR quantification methods involved. From the studies, miRNeasy Serum/Plasma kit (Qiagen) is found to be superior to other commercial kits in quality, quantity and extraction efficiency of circulating miRNAs in plasma. However modified miRNeasy Serum/Plasma kit (Qiagen) with in indicated time as per kit protocol is found to be more efficient over available commercial kits.

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