Related Experimental Study On Changes Of Intestinal Microflora In Colorectal Cancer Patients By Real-time Fluorescence Quantitative PCR

Objective:To study the changes in intestinal microflora between colorectal cancer patients and healthy people, To research specific intestinal micro-ecology background and the relationship of intestinal microflora in preoperative and postoperative patients of colorectal cancer, reveal the role and significance. Through the amount of intestinal microflora' change, screened to identify colorectal cancer at high risk, by dietary intervention and related inspection line, to early diagnosis and therapeutic. According to patients established specific changes in intestinal micro-ecology, for colorectal cancer prevention and treatment provides a new perspective.Methods:Collected the feces of preoperative and postoperative colorectal cancer patients (n=50)and healthy volunteers(n=50). According bacteria 16S rRNA gene sequence of Bifidobacterium, Lactobacillus, Escherichia coli, B.thetaiotaomicron Fusobacterium designed specific primers; According ddl gene sequence of Enterococcus faecalis and feces Enterococcu designed specific primers; According to cfxA gene sequence of Bacteroides designed specific primers; According to cfiA gene sequence of B.fragilis designed specific primers; According to mob gene sequence of B.uniformis designed specific primers; According to rpoB gene sequence of B.thetaiotaomicron,B.ovatus,B.distasoni and B.vulgatus designed specific primers; According to rrnA-16S gene sequence of Clostridium designed specific primers; According to BoNT gene sequence of C.botulinum designed specific primers; According to swp84 gene sequence of C. difficile designed specific primers; Bacterial genome DNA extracted from fecal samples were quantified by real-time fluorescence quantitative PCR to analyze bacterial amounts.Results:In normal control group and the level of Bifidobacterium spp. (9.15±0.73; 4.32±0.45; 1.47±0.28)and Lactobacillus group(7.35±0.47; 5.56±0.22; 3.52±0.36)in colorectal cancer group were significantly lower than those of healthy controls (P<0.05), whereas Escherichia coli (4.78±0.42; 5.72±0.48; 4.85±0.37) Enterococcus faecalis(4.85±0.54; 8.30±0.42; 2.48±0.44)and feces Enterococcus (5.23±0.53; 5.64±0.26; 1.55±0.29), Bacteroides(8.66±0.67; 9.75±0.87; 6.32±0.47),Fusobacterium (7.64±0.45; 10.1±0.55; 6.42±0.58),Clostridium (6.17±0.37; 6.36±0.58; 5.28±0.39),B.fragilis (3.42±0.39; 6.15±0.45; 3.25±0.58),B.uniformis(5.69±0.95; 8.70±2.18; 5.11±1.49),B.thetaiotaomicron(5.01±0.61; 9.41±2.34; 4.47±0.41),B.ovatus(4.7±1.03; 9.07±1.75; 5.22±0.49),B.distasonis(4.21±0.71; 9.53±2.46; 4.58±0.39)and B.vulgatus(2.92±1.30; 8.48±0.32; 3.80±0.87),F. necrosis(2.25±0.46;7.34±0.56; 3.58±0.45),C.botulinum (2.64±0.56; 5.39±0.43; 1.47±1.02) and C. difficile(1.65±0.51; 3.56±0.47; 1.44±0.35) were significantly higher than those of healthy controls(P<0.05), the postoperative experimental group were significantly lower than preoperative experimental group (P<0.05).Conclusions:1,Colorectal surgery treatment can affect the intestinal flora and reveal the role of pathogenesis in colorectal cancer.2,Real-time fluorescence quantitative PCR method successfully overcome the traditional bacterial culture difficulties, time-consuming, low sensitivity, inaccurate quantitative.3,Real-time fluorescence quantitative PCR method which made the leap from qualitative to quantitative and had the advantages of specific, sensitive, reproducible, accurate, fast, closed reaction, was widely used in colorectal cancer patients intestinal related moleculesmicrobial research.