A New Biomarker of Fecal Bacteria for Non-Invasive Diagnosis of Colorectal Cancer

BackgroundThe intestinal flora is correlated with the occurrence of colorectal cancer. We evaluate a new predictive model for the non-invasive diagnosis of colorectal cancer based on intestinal flora to verify the clinical application prospects of the intestinal flora as a new biomarker in non-invasive screening of colorectal cancer.MethodsSubjects from two independent Asian cohorts (cohort I, consisting of 206 colorectal cancer and 112 healthy subjects; cohort II, consisting of 67 colorectal cancer and 54 healthy subjects) were included. A probe-based duplex quantitative PCR (qPCR) determination was established for the quantitative determination of candidate bacterial markers.ResultsWe screened through the gutMEGA database to identify potential non-invasive biomarkers for colorectal cancer, including Prevotella copri (Pc), Gemella morbillorum (Gm), Parvimonas micra (Pm), Cetobacterium somerae (Cs), and Pasteurella stomatis (Ps). A predictive model with good sensitivity and specificity was established as a new diagnostic tool for colorectal cancer. Under the best cutoff value that maximizes the sum of sensitivity and specificity, Gm and Pm had better specificity and sensitivity than other target bacteria. The combined detection model of five kinds of bacteria showed better diagnostic ability than Gm or Pm alone (AUC = 0.861, P < 0.001). These findings were further confirmed in the independent cohort II. Particularly, the combination of bacterial markers and fecal immunochemical test (FIT) improved the diagnostic ability of the five bacteria (sensitivity 67.96%, specificity 89.29%) for patients with colorectal cancer.ConclusionFecal-based colorectal cancer-related bacteria can be used as new non-invasive diagnostic biomarkers of colorectal cancer. Simultaneously, the molecular biomarkers in fecal samples are similar to FIT, have the applicability in combination with other detection methods, which is expected to improve the sensitivity of diagnosis for colorectal cancer, and have a promising prospect of clinical application.

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Expression of tumor pyruvate kinase M2 isoform in plasma and stool of patients with colorectal cancer or adenomatous polyps

10.21203/rs.2.16791/v1 ◽

2019 ◽

Author(s):

Farideh Rigi ◽

Aliakbar Jannatabad ◽

Azra Izanloo ◽

Reza Roshanravan ◽

Hamid Reza Hashemian ◽

...

Keyword(s):

Colorectal Cancer ◽

Pyruvate Kinase ◽

Sensitivity And Specificity ◽

Adenomatous Polyps ◽

Plasma Samples ◽

Proliferating Cells ◽

Pyruvate Kinase M2 ◽

Specificity And Sensitivity ◽

Tumor Group ◽

Stool Samples

Abstract Background: Tumor pyruvate kinase M2 isoform (tM2-PK), which is an isoform of PK-glycolytic enzyme and appears on the surface of cancerous proliferating cells, has been used as a diagnostic biomarker for colorectal cancer (CRC). The aim of this study was to evaluate the tM2-PK measurement test for the diagnosis of CRCs and adenomatous polyps in plasma and stool samples in an Iranian population. Methods: In this prospective study, a total of 226 stool and 178 plasma samples were received from patients referred to colonoscopy units. tM2-PK enzyme was measured using two separate ScheBo-Biotech-AG ELISA kits for stool and plasma samples. Results: At the cut-off value of 4 U/ml, in tumor group, the sensitivity of fecal tM2-PK test was 100% and the specificity was 68%, and in polyp group, the sensitivity and specificity were 87% and 68%, respectively. At the cut-off value of 15 U/ml in tumor group, the sensitivity of plasma tM2-PK test was 98% and specificity was 74% and in polyp group the sensitivity and specificity were 98% and 74%, respectively. Based on our results, a cut-off range of 4.8-8 U/ml and >8 U/ml could be used to detect polyp and tumor in stool samples, respectively. Similarly, a cut-off range of 19-25 U/ml and >25 U/ml is recommended in plasma samples for polyp and tumor detection, respectively. Conclusions: This study revealed a high specificity and sensitivity of tM2-PK test for stool and plasma samples in patients with CRC and polyps suggesting it as a non-invasive assay to diagnose CRC and adenomatous polyps.

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Circulating tumor cell count from a blood sample for colorectal cancer (CRC) prevention: A 627-patient prospective study.

Journal of Clinical Oncology ◽

10.1200/jco.2019.37.4_suppl.485 ◽

2019 ◽

Vol 37 (4_suppl) ◽

pp. 485-485 ◽

Cited By ~ 2

Author(s):

Wen-Sy Tsai ◽

Drew Watson ◽

Ying Chang ◽

Ben Hsieh ◽

Hung-Jen Shao ◽

...

Keyword(s):

Colorectal Cancer ◽

Logistic Regression ◽

Prospective Study ◽

Compliance Rate ◽

Screening Method ◽

Unmet Need ◽

Detection Methods ◽

P Value ◽

Non Invasive ◽

Adenoma Detection

485 Background: Up to 25% of patients with adenomas progress to having colorectal cancer. If detected early, adenomas can be removed with a diagnostic colonoscopy procedure, preventing cancer. Invasive colonoscopy is the only screening method with the sensitivity to accurately detect adenomas, but has a low compliance rate of 38% for screening. Available non-invasive tests (including stool-based multi-analyte tests) have very limited sensitivity for adenomas. Hence, there is an unmet need for a non-invasive test for adenoma detection. Methods: IRB-approved prospective study was conducted in 627 subjects 50 years or older- recommended for routine CRC screening- 405 subjects had adenoma or CRC, confirmed by colonoscopy with tumor biopsy. Two mL peripheral blood was processed using the CellMax biomimetic platform (CMx), which uses a microfluidic biochip to enumerate circulating tumor cells (CTCs). Nominal logistic regression was used to assess performance while proportional odds logistic regression and Cuzick’s trend test were used to determine association of CTC counts with cancer stage. Results: An increase in CTC count was significantly correlated with an increase in disease burden (Cuzick’s Test p-value < 0.0001). Furthermore, there was a significant association between CTC counts and stages of adenoma-carcinoma progression (Likelihood ratio p-value < 0.0001). The CTC enumeration was able to differentiate between healthy and diseased patients (adenoma + cancer). Conclusions: To the best of our knowledge, these are the first reported results for a blood test that has high accuracy for adenoma detection, and truly enables colorectal cancer prevention. This test can be administered in the primary care setting and drive high compliance.[Table: see text]

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Urinary Measurement of Epigenetic DNA Modifications: A Non-Invasive Assessment of the Whole-Body Epigenetic Status in Healthy Subjects and Colorectal Cancer Patients

ChemistryOpen ◽

10.1002/open.201600103 ◽

2016 ◽

Vol 5 (6) ◽

pp. 550-553 ◽

Cited By ~ 4

Author(s):

Rafal Rozalski ◽

Daniel Gackowski ◽

Agnieszka Siomek-Gorecka ◽

Zbigniew Banaszkiewicz ◽

Ryszard Olinski

Keyword(s):

Colorectal Cancer ◽

Cancer Patients ◽

Healthy Subjects ◽

Whole Body ◽

Non Invasive ◽

Dna Modifications ◽

Colorectal Cancer Patients

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Elucidating the Gut Microbiome of Colorectal Cancer: 40 Fecal Bacteria as Non-invasive Biomarkers

10.21203/rs.3.rs-113328/v1 ◽

2020 ◽

Author(s):

Biao Yuan ◽

SiPing Ma ◽

Qingkai Meng ◽

Tao Du ◽

Yueyan Zhu ◽

...

Keyword(s):

Microbial Community ◽

Early Diagnosis ◽

Sensitivity And Specificity ◽

Gut Microbiome ◽

Microbial Community Analysis ◽

Support Vector ◽

Healthy Controls ◽

Microbial Composition ◽

Non Invasive ◽

Specificity And Sensitivity

Abstract Background: Colorectal adenocarcinoma (CRC) ranks one of the 5 most lethal malignant tumors both in China and worldwide. Emerging evidences have revealed the importance of gut microbiome on CRC, thus microbial community could be termed as a potential screen for early diagnosis. Importantly, compared with the whole microbial community analysis, few numbers of bacteria genus as non-invasive biomarkers with high sensitivity and specificity causing less cost would benefit more in clinical. Methods: Here we analyzed the gut microbiome from 226 CRC patients and 156 healthy people by 16s rRNA sequencing. We analyzed the microbiome diversity between CRC patients and healthy controls. We used ExtraTrees classifier to screening the biomarkers and took SVM (Support Vector Machine) model to test the specificity and sensitivity of our biomarkers. Results: Compared with the healthy gut, the microbial composition are divergent in CRC, especially the increase of some bacteria related to CRC and the decrease of some healthy bacteria. 40 bacteria genus exhibiting high weight for the healthy and CRC microbiome classification were screened as biomarkers for CRC. In addition, the combination of 40 biomarkers and FOBT showed an outstanding sensitivity and specificity for discrimination CRC patients from healthy controls. Conclusion: The method could be used as a non-invasive method for CRC early diagnosis.

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Advances in early detection of colorectal cancer: A focus on non-invasive biomarkers

Current Drug Targets ◽

10.2174/1389450122666210303100048 ◽

2021 ◽

Vol 22 ◽

Author(s):

Nadia El Kadmiri

Keyword(s):

Colorectal Cancer ◽

Sensitivity And Specificity ◽

Secondary Outcome ◽

Diagnostic Biomarkers ◽

Non Invasive ◽

Biomarker Analysis ◽

Genomic Markers ◽

Bibliographic Search ◽

Noninvasive Tests

Background: Colorectal cancer (CRC) is the third most commonly diagnosed cancer Worldwide. Currently, colonoscopy remains the gold standard diagnostic test for CRC detection. Nonetheless, this technique is invasive and expensive. Remarkable ongoing strategies are focusing on development affordable methods to diagnosis at earlier stages and surveille CRC. The introduction of suitable noninvasive, sensitive and specified diagnostic tests for early CRC detection by employing biomarker analysis seems to be a fundamental need to reduce the numbers of unnecessary colonoscopies. In this Review, we provide an overview of single- and multi-panel biomarkers (Genomic markers, transcriptome markers, proteomic markers, inflammatory markers and microbiome markers) encompassing noninvasive tests in blood and stool for early CRC detection. Methods: A bibliographic search using PubMed/Medline, Web of Science and EBSCOhost databases was performed to find relevant published studies over the last 6 years. Forty-three pertinent studies were included in this review. Results: The primary outcome highlights the sensitivity and specificity of single diagnostic biomarkers studied in blood or stool. The secondary outcome reveals the sensitivity and specificity of panel -biomarkers (combinations) in blood or stool. While some markers show better performance, other are not suitable for screening purposes. Conclusion: It remains to adjust experimental and analytical tests that can interfere à robust results to replace or supplement those currently markers in use. Even so, robust verification and validation with large clinical cohorts are needed to successful noninvasive tests that can fulfill the role of colonoscopy.

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Cell-free circulating DNA as a promising marker of colorectal cancer detection and progression

Journal of Clinical Oncology ◽

10.1200/jco.2009.27.15_suppl.11059 ◽

2009 ◽

Vol 27 (15_suppl) ◽

pp. 11059-11059

Author(s):

S. Pucciarelli ◽

M. Enzo ◽

M. Agostini ◽

S. Pizzini ◽

P. Del Bianco ◽

...

Keyword(s):

Colorectal Cancer ◽

Healthy Subjects ◽

P Value ◽

Circulating Dna ◽

Plasma Samples ◽

Alu Repeats ◽

Non Invasive ◽

Total Dna ◽

Free Circulating Dna ◽

Normal Controls

11059 Background: Since the pathologic stage is the most powerful prognostic factor for colorectal cancer (CRC), there is a strong need of non-invasive methods for early detection. Cell-free circulating DNA (cfDNA) released from cancer cells varies in size. It has been suggested that cfDNA (ALU repeats of 115 bp, representative of total DNA; ALU repeats of 247 DNA, representative of tumor DNA) may be associated with presence of tumor. Aim of this study was then to investigate whether the cfDNA may have a role as marker of CRC detection and progression. Methods: cfDNA was extracted from plasma samples from 136 patients with primary CRC at different stages [median age 64 yrs; male/female 78/58; stages I-II, 61; stages III-IV, 75], and from 24 patients with adenomas [median age 67 yrs; male/female 17/7)] and from 55 clean-colon healthy subjects [median age 56 yrs; male/female 13/43). cfDNA was assessed by quantitative real-time PCR (qPCR) of ALU repeats with 2 sets of primers (115 and 247 bp) amplifying different lengths of DNA. The levels of cfDNA (ALU-115, ALU-247) of CRC patients (stages I-II and stages III-IV) were compared with those of healthy subjects and patients with adenoma. Results: The median concentrations of total cfDNA (ALU115) in the plasma samples from patients with stages III-IV and stages I-II CRC, adenoma and normal controls were 52,4, 11.9; 1.9, and 1.7 ng/ml, respectively (p<.0001). The corresponding figures for tumor-related cfDNA (ALU247) were 48.8, 4.7, 2.2, and 0.7 ng/ml, respectively. (p<.0001). With a cut-off of 4.86 ng/ml, total DNA (ALU115) showed a sensitivity of 78.52 (95% CI 70.6–85.1) and a specificity of 86.08 (95% CI 76.4–92.8) in distinguishing patients with CRC from non-CRC [AUC: 0.860 (95% CI 0.81–0,90), p-value=.0001]. With a cut-off of 3.04, cfDNA tumor-related (ALU247) showed a sensitivity of 77.94 (95% CI 70.0–84.6) and a specificity of 82.28 (95% CI 72.1–90.0) in distinguishing patients with CRC from non-CRC [AUC: 0.864 (95% CI 0.81–0,91), p-value=.0001]. Conclusions: Both ALU115 and ALU 247 fragments of circulating cfDNA seem promising non-invasive molecular markers of detection and progression of CRC. The findings of the current study require to be confirmed on larger cohorts of patients with CRC and colonic adenoma. No significant financial relationships to disclose.

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Investigations of a novel diagnostic technology for colorectal cancer.

Journal of Clinical Oncology ◽

10.1200/jco.2017.35.4_suppl.564 ◽

2017 ◽

Vol 35 (4_suppl) ◽

pp. 564-564

Author(s):

Hongmei Tao ◽

Xuedong Du ◽

Shijin Ding ◽

Xing Tang ◽

Da Lou ◽

...

Keyword(s):

Colorectal Cancer ◽

Sensitivity And Specificity ◽

Large Population ◽

Diagnostic Methods ◽

Control Group ◽

P Value ◽

Blood Samples ◽

Non Invasive ◽

Cancer Group ◽

Diagnostic Technology

564 Background: Currently endoscopy examination takes an important role in the diagnosis of colorectal cancer, and there are no other clinically viable non-invasive diagnostic methods. In this investigation, a newly developed, novel IVD cancer diagnostic technology named Cancer Differentiation Analysis (CDA) Technology was investigated for colorectal cancer diagnosis. CDA is a new technology using multi-level and multi-parameter information which measures information relating to both protein fragments and cellular signals in blood samples in one single test. Methods: Peripheral blood was drawn in EDTA tubes for CDA tests. Intravenous blood samples from individuals with colorectal cancer (n = 193), as well as control samples (n = 705) were collected. Cancer group has been clinically diagnosed, and individuals in control group have been confirmed by physical examinations free of cancer. All blood samples were tested using a CDA device, and collected data were analyzed in the SPSS Packages. Results: Data analysis (using T test) showed a significant statistical difference with P value < 0.05 between colorectal cancer group and control group. Details of CDA test results were given in Table 1. Based on initial data, the area under curve (AUC), sensitivity and specificity of CDA technology for colorectal cancer were determined to be 0.861, 76.7% and 76.7% (cut-off value was set at a level at which sensitivity and specificity are comparable), respectively. Conclusions: CDA technology is able to statistically distinguish control group from colorectal cancer group with reasonably high sensitivity and specificity (both above 75%). As a non-invasive and potentially cost effective method capable of large population screening, CDA technology could be a very promising approach for the screening and diagnosis of colorectal cancer. [Table: see text]

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Evaulation of miRNAs as a Non-invasive Blood-Based Marker for Detection of Colorectal Cancer

10.21203/rs.3.rs-634213/v1 ◽

2021 ◽

Author(s):

Mohamed Fakhry ◽

Huda Mohamed ◽

Mona Mohamed ◽

Sara Adel ◽

Mohammed Tag-Adeen ◽

...

Keyword(s):

Gene Expression ◽

Colorectal Cancer ◽

Rectal Cancer ◽

Healthy Subjects ◽

Area Under Curve ◽

Gastrointestinal Endoscopy ◽

Control Group ◽

Group Level ◽

Non Invasive ◽

American Society

Abstract Backgrounds: Although Colorectal cancer accounts for over 10% of all cancer incidence, it is highly preventable and can be detected at a stage when there are often no symptoms. The American Society for Gastrointestinal Endoscopy encourages everyone over 50 to be screened for colorectal cancer, but many should start much earlier due to health and family history. Minimally invasive tests will be superior overtime taking into consideration availability, costs, convenience, and patient–clinicians preferences. Methods: 50 patients with known colorectal cancer (CRC) were enrolled based on colonoscopic and histopathological findings and 25 healthy subjects were enrolled as control group. Level of microRNA-21 gene expression was measured in both groups. Results: Level of microRNA-21 gene expression was significantly higher among CRC group in comparison to control group (2.76 ± 0.56 vs. 0.71 ± 0.16 (u/l); P< 0.001). A cut off point > 1.09 u/l, microRNA-21 gene expression had 96% sensitivity and 95% specificity with area under curve was 0.95 for detection of colo-rectal cancer. Level of microRNA-21 gene expression was significantly higher among CRC group in comparison to control group and its level was increasing with those with metastasis.Conclusion: Level of microRNA-21 gene expression is useful biomarker for discriminating CRC patients from healthy people.

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Rearrangements of Blood and Tissue Fatty Acid Profile in Colorectal Cancer - Molecular Mechanism and Diagnostic Potential

Frontiers in Oncology ◽

10.3389/fonc.2021.689701 ◽

2021 ◽

Vol 11 ◽

Author(s):

Adriana Mika ◽

Katarzyna Duzowska ◽

Lukasz P. Halinski ◽

Alicja Pakiet ◽

Aleksandra Czumaj ◽

...

Keyword(s):

Colorectal Cancer ◽

Fatty Acid ◽

Healthy Subjects ◽

Roc Analysis ◽

Normal Mucosa ◽

Cancer Tissue ◽

Normal Colon Mucosa ◽

Non Invasive ◽

Diagnostic Potential ◽

Ht 29

Colorectal cancer (CRC) is often diagnosed at an advanced stage due to the invasiveness of colonoscopy; thus, non-invasive CRC diagnostics are desirable. CRC is associated with lipid alterations. We aimed to verify whether fatty acid (FA) profiles in CRC patients may serve as a potential diagnostic tool for CRC diagnosis. FA profiles were assayed by GC-MS in cancer tissue, paired normal mucosa and serum from CRC patients and healthy controls. The levels of very long FAs – VLCFAs (26:0, 28:0 and 26:1) were the most highly increased FAs in cancer tissue compared to normal colon mucosa. Moreover, these FA were present in serum of CRC patients, they were absent in the serum of healthy subjects, or present in only trace amounts. To verify if cancer cells are the source of small amounts of these VLCFAs in the serum of patients we performed experiment in HT-29 CRC cells, which proved that CRC cells can produce and release VLCFAs into the blood. Most importantly, we defined a panel of FAs that may be assayed in a single analysis that definitely distinguishes CRC patients and healthy subjects, which was confirmed by PLS-DA and multivariate ROC analysis (AUC = 0.985). This study shows that selected FA panel may serve as a diagnostic marker for CRC.

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Performance of molecular methods for the detection of Salmonella in human stool specimens

Wellcome Open Research ◽

10.12688/wellcomeopenres.16305.1 ◽

2020 ◽

Vol 5 ◽

pp. 237

Author(s):

Angeziwa Chunga Chirambo ◽

Tonney S. Nyirenda ◽

Ndaru Jambo ◽

Chisomo Msefula ◽

Arox Kamng'ona ◽

...

Keyword(s):

Multiplex Pcr ◽

Sensitivity And Specificity ◽

Stool Culture ◽

Test Methods ◽

Detection Methods ◽

Molecular Diagnostic ◽

Specificity And Sensitivity ◽

Stool Specimens ◽

Low Sensitivity ◽

Pcr Targeting

Background: The relationship between asymptomatic Salmonella exposure within the gastrointestinal tract and Salmonella bacteraemia is poorly understood, in part due to the low sensitivity of stool culture, and the lack of validated molecular diagnostic tests for the detection of Salmonella in stool. The study aimed to determine a reliable molecular diagnostic test for Salmonella in stool specimens. Methods: We optimized an in-house monoplex real time polymerase chain reaction (PCR) for the detection of Salmonella TTR and InvA genes in stool by including a selenite broth pre-culture step for Salmonella before DNA extraction, and validated their specificity against other local common pathogens. Then we assessed their performance against a well-validated multiplex PCR targeting the same TTR and InvA genes, and against stool culture using clinical stool specimens collected from a cohort of 50 asymptomatic healthy Malawian children that were sampled at 1-month intervals over a period of 12 months. We employed a latent Markov model to estimate the specificities and sensitivities of PCR methods. Results: TTR and InvA primers were both able to detect all the different Salmonella serovars tested, and had superior limits of detection if DNA was extracted after selenite pre-culture. TTR sensitivity and specificity for monoplex-PCR were (99.53%, 95.46%) and for multiplex-PCR (90.30%, 99.30%) respectively. InvA specificity and specificity for using monoplex-PCR was (95.06%, 90.31%) and multiplex-PCRs (89.41%, 98.00%) respectively. Sensitivity and specificity for standard stool culture were 62.88% and 99.99% respectively. Culture showed the highest PPV (99.73%) and mono-TTR had the highest NPV (99.67%). Conclusion: Test methods demonstrated high concordance although stool culture and monoplexed TTR primers had superior specificity and sensitivity respectively. The use of selenite pre-enrichment step increased Salmonella detection rate. Taken together, molecular detection methods used here could be used to reveal the true extent of both asymptomatic and symptomatic Salmonella exposure events.

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