XÂY DỰNG ĐỐI CHỨNG CHUẨN CHO KỸ THUẬT MULTIPLEX qPCR XÁC ĐỊNH BACTEROIDES FRAGILIS, FUSOBACTERIUM NUCLEATUM, AKKERMANSIA MUCINIPHILA VÀ ESCHERICHIA COLI Ở BỆNH NHÂN UNG THƯ ĐẠI TRỰC TRÀNG

Abstract

Disorders of gut microbiota may alter physiological processes resulting in various diseases. The presence of toxigenic Bacteroides fragilis (ETBF), Fusobacterium nucleatum (Fn), Akkermansia muciniphila (Am) and Escherichia coli (E.coli), have been implicated as agents associated with onset and progression. of colorectal cancer through the secretion of toxins or oncogen activators.

As such, our study aimed to develop a standard calibrator; develop a simple, rapid technique to simultaneously detect ETBF, Fn, Am, E.coli in tumors of CRC patients using multiplex qPCR. Specific primers and probes were designed based on the highly conserved regions  of  ETBF, Fn, Am, E.coli, built a standard calibrator and optimized multiplex qPCR conditions. As a result, we have created four recombinant plasmids.The sensitivity of the technique reaches 10 copies, with correlation coefficient R² >0.99; E > 93% and can be used as a standard control for analyzing the specimens from patients. At the same time, detected positivity for Fn, ETBF, and NTBF in 65%, 66%, 62%, and 36%. of the samples respectively (n = 100). These preliminary findings suggest that disruptions in the gut microbiota have the potential to serve as a biomarker for the early detection of CRC.