Abstract
The SEPT9 gene methylation assay is the first FDA-approved blood assay for colorectal cancer (CRC) screening. Fecal immunochemical test (FIT), FIT-DNA test and CEA assay are also in vitro diagnostic (IVD) tests used in CRC screening. This meta-analysis aims to review the SEPT9 assay performance and compare it with other IVD CRC screening tests. By searching the Ovid MEDLINE, EMBASE, CBMdisc and CJFD database, 25 out of 180 studies were identified to report the SEPT9 assay performance. 2613 CRC cases and 6030 controls were included, and sensitivity and specificity were used to evaluate its performance at various algorithms. 1/3 algorithm exhibited the best sensitivity while 2/3 and 1/1 algorithm exhibited the best balance between sensitivity and specificity. The performance of the blood SEPT9 assay is superior to that of the serum protein markers and the FIT test in symptomatic population, while appeared to be less potent than FIT and FIT-DNA tests in asymptomatic population. In conclusion, 1/3 algorithm is recommended for CRC screening, and 2/3 or 1/1 algorithms are suitable for early detection for diagnostic purpose. The SEPT9 assay exhibited better performance in symptomatic population than in asymptomatic population.
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Introduction
Colorectal cancer (CRC) has become the 3rd leading cause of new cancer cases in the world. The prevention of CRC should aim at early detection, which can be achieved by regular screening. Statistics from U.S. Preventive Services Task Force (USPSTF) shows that approximately 60% CRC deaths could be avoided if a regular periodic screening was carried out each year, and the average five-year survival rate could be increased from 46 to 73%1. Therefore, effective CRC early screening methods can prolong patients’ lives and reduce mortality. The latest recommendation statement from the USPSTF listed the stool-based gFOBT (guaiac-based fecal occult blood test), FIT (fecal immunochemical test), FIT-DNA (multitargeted stool DNA) tests, the blood-based SEPT9 gene methylation assay and the direct visualization tests (including colonoscopy, CT colonoscopy and sigmoidoscopy) as the current CRC screening strategies2.
The blood-based SEPT9 gene methylation assay aim at detecting the aberrant methylation at the promoter region of the SEPT9 gene DNA released from CRC cells into the peripheral blood (i.e. the circulating tumor DNA, or ctDNA)3, 4. The development of the test is based on a key theory indicating that the detection of the SEPT9 gene aberrant methylation reflects the existence of CRC. The CpG island 3 at the promoter region of the SEPT9 gene V2 transcript is hypermethylated, and DNA of the gene is released into the peripheral circulating blood from necrotic and apoptotic cancer cells during CRC carcinogenesis5. The risk of CRC can be determined by detecting the degree of DNA methylation of the specific promoter region of the SEPT9 gene in the peripheral blood6.
There are 25 independent studies performed so far to investigate the performance of the SEPT9 gene methylation assay in CRC detection, in which most studies were case-control or cohort studies, while only one randomized multi-center screening study and two opportunistic screening studies were performed to investigate its performance in average-risk population and high-risk population, respectively7, 9,10,11. However, the algorithm (1/3 algorithm) used in the screening study was also applied in some other studies, and their performance was therefore comparable to each other. Other algorithms, including 2/3, 1/1 and 1/2, were also used in some studies. There is no consensus on which algorithm is superior in various occasions. Moreover, as FIT and FIT-DNA tests are also CRC screening assays currently used, it would be useful to compare their performance with the SEPT9 assay.
In this systematic review and meta-analysis, we investigated the performance of SEPT9 assay with specific focuses on test sensitivity and specificity at various algorithms. The overall test performance and the stage-dependent sensitivity with different algorithms were compared. We also compared the assay performance with that of the serum protein tumor markers, FIT and FIT-DNA tests. The objective of this study is to identify the best algorithm for various scenarios, and to highlight the pros and cons of the SEPT9 assay in CRC screening.
Results
Eligibility of studies and literature characteristics included in the meta-analysis
Twenty-five studies investigating the performance of the SEPT9 assay at various settings are obtained from the screening through PRISMA flow diagram (Fig. 1) and are listed in Table 1. These studies were accessed using the QUADAS system8 and the results were shown in Supplementary Figure 1. Several parameters, including sensitivity, specificity, positive likelihood ratio (PLR), negative likelihood ratio (NLR), odds ratio (OR), algorithm, and kit used in each study, were compared in Table 1. In the 25 reports, 22of them were cohort or case-control studies, while the PRESEPT study is the only one carried out so far in the screening background in average-risk population7, 9, and the RESEPT study and a recent report by Song et al. performed the opportunistic screening in high-risk population10, 11. 3 studies adopted the 1/3 algorithm alone9, 12, 13, 3 studies adopted the 1/2 algorithm alone7, 14, 15, 6 studies adopted the 1/1 algorithm alone3, 10, 16,17,18,19, and 7 studies adopted the 2/3 algorithm alone4, 20,21,22,23,24,25, while 6 studies tested both 1/3 and 2/3 algorithm6, 11, 26,27,28,29.
The blood SEPT9 gene methylation exhibits adequate sensitivity and specificity in CRC detection and screening
Table 1 summarized the sensitivity, specificity, PLR, NLR, OR, algorithm and kits used in each study. The sensitivity and specificity of the cohort or case-control study were affected by study design, population, selection of cases, choice of kits and algorithm, etc. Generally speaking, the sensitivity of these studies ranged from 48.2% to 95.6%, with the specificity ranged from 79.1% to 99.1% (Table 1). The latest commercialized SEPT9 assay, the Epi proColon 2.0, exhibited a higher sensitivity at 71.1–95.6%, and maintained high specificity at 81.5 to 99% (Table 1). The effect of different algorithm can be observed in studies with multiple algorithms applied, and it will be discussed further in this article.
The PLR of the 25 studies ranged from 2.87 to 73.00, exhibiting a high ratio between true positive and false positive rate and suggesting a high probability of true positive when a test result is positive. The NLR of the 25 studies ranged from 0.05 to 0.57, exhibiting a high ratio between false negative and true negative rate and suggesting a high probability true negative when a test result is negative. The OR of the 25 studies ranged from 5.67 to 349.63, indicating that the SEPT9 methylation is a high-risk factor and has diagnostic significance for CRC. PLR, NLR and OR exhibited a big variation among all studies. This may be due to different design of studies, since various inclusion of CRC cases, non-CRC colonic diseases, and normal controls in case-control, cohort or screening studies can greatly affect the positive and negative rate. This may also be due to the difference in kit performance, as those early stage research kits and commercialized kits exhibited lower detection capability than later improved kits.
Currently, the PRESEPT study in the only screening study performed in average-risk population from 50 to 75 years old. The sensitivity reported (48.2%) using 1/2 algorithm was apparently lower than those reported in previous cohort or case-control studies. In a later report by Potter and colleagues9, triplicate PCRs were performed using samples from the same study. The sensitivity increased to 68.2% and the specificity decreased to 80.0%. The US FDA approved the Epi proColon, the commercialized SEPT9 assay, based on the data from the PRESEPT study with 1/3 algorithm7, 9. Apart from the screening in average-risk population, the SEPT9 assay was also used in the opportunistic screening for high-risk population. In one study performed in four northern Chinese hospitals (RESEPT study) using the SensiColon assay, the SEPT9 assay exhibited a sensitivity of 76.6% with a specificity of 95.9% at a total positive rate of 25.8%10. In another recent opportunistic screening study using the Epi proColon 2.0 CE kit, the SEPT9 assay exhibited a sensitivity of 75.1% with a specificity of 95.1%11.
The choice of algorithm affects the performance of the SEPT9 assay
Table 1 lists the four algorithms currently used in studies of SEPT9 assay. The positive test results were determined by one positive count out of three PCRs (1/3 algorithm), one positive count out of two PCRs (1/2 algorithm), two positive counts out of three PCRs (2/3 algorithm), or one positive count out of one PCR (1/1 algorithm). It can be clearly seen that a majority of studies performed 1/3 or 2/3 algorithm, while 1/2 and 1/1 algorithm were also used for some studies.
In order to compare the performance of SEPT9 assay at various algorithms, study data from each algorithm were pooled and meta-analyzed. As shown in Fig. 2, 1/3 algorithm exhibited the best sensitivity (0.78) with lowest specificity (0.84) among all algorithms (Fig. 2A), while 2/3 algorithm exhibited the highest specificity (0.96) (Fig. 2B). The sensitivity and specificity of 2/3 and 1/1 algorithm (Fig. 2B and D) were very similar (sensitivity: 0.73 vs 0.74, specificity: 0.96 vs 0.94). 1/2 algorithm (Fig. 2C) exhibited the lowest sensitivity (0.59) with satisfactory specificity (0.91). The area under the curve (AUC) showed very similar values for 1/3, 2/3 and 1/2 algorithm, while the AUC for 1/1 algorithm appeared to be slightly lower than others. It can be observed that the 1/3 algorithm provides the best sensitivity at the price of lower specificity, while the 2/3 algorithm provides the best balance between sensitivity and specificity.
SEPT9 assay is effective for the detection of early-stage CRC
Although SEPT9 assay exhibited overall high sensitivity and specificity in detecting CRC, the ability of detecting early-stage CRC is more important than later stages of CRC, since early detection can ensure early intervention to enhance the cure rate and reduce mortality. Stage I and II are regarded as early stages in this article. Table 2 summarizes the stage-related detection rate of all available studies and the pooled data categorized by the algorithm used in these studies. The data in Table 2 is plotted in Fig. 3 to compare the detection rate at each stage (Fig. 3A) and the effect of algorithm on stage-dependent detection rate (Fig. 3B). It can be seen from Table 2 and Fig. 3 that there is a clear trend in which the detection rate increases with the escalation of clinical stage, indicating that the level of SEPT9 methylation had a correlation with the degree of malignancy, regardless of the algorithm. Furthermore, the algorithm exhibited a clear effect on the sensitivity for every stage (Fig. 3B). The sensitivity with various algorithm can be ranked as 1/3 > 2/3 > 1/1 > 1/2 from highest to lowest.
SEPT9 assay generally exhibited the highest detection rate for all stages of CRC with 1/3 algorithm (Fig. 3B). The detection rate for stage I and II reached 59.6% and 85.7%, respectively (Table 2), representing a very high detection rate for early-stage CRC among current in-vitro diagnostic methods. The SEPT9 assay with 2/3 or 1/1 algorithm detected approximately half of stage I and 70% of stage II CRC, respectively, which was also satisfactory for early CRC detection (Table 2, Fig. 3B). Apparently, for CRC screening aiming at early-stage cancer detection, algorithms with high sensitivity should be adopted. However, false positive detection will also increase with higher sensitivity algorithms. These data clearly show that the SEPT9 assay is effective for early stage CRC detection.
The Performance of the SEPT9 assay is superior to CEA and the FIT tests in the screening of symptomatic population
In order to further evaluate the performance of the SEPT9 assay in CRC screening and diagnosis, the sensitivity and specificity of the assay was compared with the current clinically used serum markers (CEA, CA50, CA242, CA724 and CA199)32 in the screening of symptomatic population. As 1/3 algorithm was recommended by the US FDA as the interpretation method, the pooled data from 1/3 algorithm of the assay was used for comparison. The sensitivity and specificity of these tests were meta-analyzed by reviewing the study on the performance of each individual test on CRC detection. The Forest plot was made for each serum marker and the estimated sensitivity and specificity were analyzed for comparison with the SEPT9 assay (Fig. 4A and B). It can be clearly seen from Fig. 4A and B that the sensitivity of the SEPT9 assay (78%) in CRC detection was much higher than any of the five serum markers, and its specificity (84%) was at the same range as these serum markers, including the most commonly used CRC serum marker, CEA. The detailed data for the serum markers was summarized and analyzed in Supplementary Figure 2 (CEA), 3 (CA50), 4 (242), 5 (CA724) and 6 (CA199), respectively.
As FIT is a test widely used for CRC screening in symptomatic population, three studies compared the performance of FIT with that of the SEPT9 assay side by side. Table 3 summarized the sensitivity and specificity of FIT and the SEPT9 assay in the three studies. It can be seen from the pooled data that the SEPT9 assay exhibited significantly higher sensitivity than the FIT test (75.6% vs 67.1%, p < 0.05), while they showed essentially identical specificity. It appeared that the performance of the SEPT9 assay in screening of symptomatic population is better than that of the FIT test.
The SEPT9 assay is less potent than the FIT and the FIT-DNA tests in the screening of asymptomatic population
Currently, the PRESEPT study7, 9 is the only study investigated the performance of the blood SEPT9 assay in average-risk asymptomatic population, we therefore compared the data from this report with data from FIT33 and FIT-DNA test34 in the same type of population. It can be clearly seen from Fig. 5A and B that SEPT9 exhibited lower sensitivity (68.0% for the SEPT9 assay, compared with 79.0% for FIT and 92.3% for FIT-DNA test) and lower specificity (80.0% for the SEPT9 assay, compared with 94.0% for FIT and 86.6% for FIT-DNA test) than FIT or FIT-DNA test. The performance of the blood SEPT9 assay in asymptomatic population screening appeared to be lower than that of the FIT and FIT-DNA tests. However, the SEPT9 assay exhibited better compliance than FIT test. One recent study35 showed that 63% of subjects recommended for CRC screening refused colonoscopy screening. 97% of subjects refusing colonoscopy accepted a noninvasive screening test, in which 83% chose the Septin9 blood test and 15% chose FIT test. It is clear from the study that patients prefer SEPT9 test than colonoscopy and FIT35.
Discussion
It is notable that the screening sensitivity of 48.2% reported in the PRESEPT study was much lower than those reported in previous cohort or case-control studies. This can be explained from three aspects. Firstly, duplicate PCR reactions, instead of triplicate PCRs, were used in this study. The sensitivity in this study was obtained from at least one positive reaction out of two PCRs, instead of three PCRs, therefore, the chance to detect abnormally methylated SEPT9 DNA was lower than those previous studies performing three PCRs. Secondly, the study setting was different from those previous studies. The PRESEPT study aimed at screening of asymptomatic average-risk population between 50 and 75 years old. CRC patients found in the asymptomatic population are more likely to be those with early-stage CRC. As the sensitivity in early-stage CRC appeared to be lower than the overall sensitivity from all stages, the sensitivity from the asymptomatic population tends to be lower than those with symptomatic population, i.e. CRC subjects recruited from hospitals in those case-control or cohort studies. Thirdly, the reaction system in different studies varied, and this may explain why 1/1 algorithm exhibited higher performance than 1/2 algorithm. The PCR reaction from Epi proColon series product used 30 µl reaction system, however, is was reported that the latest SensiColon product used 60 µl reaction system with doubled amount of DNA template10. This allowed higher chance of methylated DNA detection, although only one PCR reaction was performed.
The PRESEPT study represents the scenario in the real life CRC screening setting and therefore has better guiding significance than case control studies. The application of three PCRs was based on the consideration of identifying as many cancer patients as possible in the screening background. It appears that the sensitivity reported in the study using 1/3 algorithm (68.2%) was much better than that obtained from 1/2 algorithm9. Therefore, 1/3 algorithm was accepted by the US FDA for the approved product.
Apart from the screening of average-risk population, the SEPT9 assay was also used in the opportunistic screening of high-risk population, in which the chance of identifying positive subjects is much higher than that in average-risk population, as this screening is commonly performed in hospital environment. Therefore, the positive rate, sensitivity and specificity are related to specific population in a screening. The SEPT9 assay exhibited high sensitivity and specificity in opportunistic screening10. These parameters appeared to be higher than those in PRESEPT study, as the case composition in this opportunistic screening was quite different from that in REPSEPT study.
A distinct feature of the SEPT9 assay is that most studies performed multiple PCRs to enhance test sensitivity. This is because SEPT9 assay is designed for detecting trace amount of methylated SEPT9 gene copies in strong background of unmethylated SEPT9 DNA, and the amount of detectable methylated SEPT9 DNA can be as low as 7.8 pg/ml, equivalent to 1–2 copies of genome DNA9. This leads to the question of interpreting the PCR data when some of the PCR reactions show positive while others show negative results. Due to the application of different algorithms in data analysis, sensitivity and specificity vary with different methods of interpretation.
The choice of algorithms is based on the specific needs in a test. Two aspects need to be considered before a method can be properly chosen. One is to exclude healthy negative population, which requires high specificity, and the other is to identify as many real patients as possible to enhance the disease detection rate, which requires high sensitivity. These two aspects are normally against each other, as they are two factors at the ends of a teeterboard. As the 1/3 algorithm showed the best sensitivity while the 2/3 algorithm showed the best specificity, the choice of 1/3 algorithm or 2/3 algorithm depends on the purpose of a study. If detecting cancer is the main purpose, such as that in a screening test, 1/3 algorithm can be used to maximize the number of patients detected. The situation of false positive subjects can be confirmed by further diagnostic method, such as colonoscopy. In contrast, if the main purpose is to exclude healthy subjects to ensure low misdiagnosis rate, 2/3 algorithm can be used, and routine screening program should be implemented to minimized the ratio of missed cancer patients.
As the SEPT9 assay was shown to detect early-stage CRC with high sensitivity, it has great advantages over serum protein markers in CRC early detection. CEA is the most commonly used markers for CRC detection, however, due to its low sensitivity for early-stage CRC, it is more widely used in post-surgery monitoring of CRC recurrence and therapeutic effects, rather than a screening marker. SEPT9 gene methylation appeared to be the best blood-based single marker for CRC screening and early detection so far.
Although the SEPT9 assay did not exhibit similar sensitivity and specificity compared with FIT and FIT-DNA in asymptomatic population screening, it is a competitive option for CRC screening and early detection, as it has been shown to have higher compliance in CRC screening than FIT and colonoscopy35. A good screening test should be one that not only has high sensitivity and specificity, but also has high uptake rate by population.
Conclusions
The SEPT9 assay is the first blood-based test aiming at ctDNA detection for CRC screening and early detection. It shows a high sensitivity and specificity in CRC screening and early detection. The choice of algorithm is based on the needs for a test. Algorithm with high sensitivity (1/3) can be used in screening, while algorithm with high specificity (2/3 or 1/1) can be used in CRC early detection for diagnostic purpose to exclude normal subjects. The SEPT9 assay exhibited better performance than protein markers and FIT test in symptomatic population, while showed lower sensitivity and specificity than the FIT and the FIT-DNA tests in asymptomatic population. However, it provides an effective option for patients who have low compliance with FIT or colonoscopy.
Methods
All literature search, selection, data extraction, study quality assessment, and data analysis were performed based on the rules, guidelines or recommendations from the preferred reporting items for systematic reviews and meta-analyses (PRISMA) and the quality assessment for diagnostic accuracy studies (QUADAS)8, 30, 31, 36, 37. Relevant softwares were used for the above process and data analysis, including Review Manager 5.2 (The Cochrane Collaboration, London, UK), Stata 14.0 (StataCorp LP, TX, USA), MetaDisc 1.4 (Unit of Clinical Biostatistics team of the Ramóny Cajal Hospital, Madrid, Spain) and PRISM 5.0 (GraphPad Software, Inc., La Jolla, CA, USA).
Search strategy
The Ovid MEDLINE,EMBASE, CBMdisc (China Biology Medicine disc) and CJFD (Chinese Journal Full—text Database) database were searched using the key words ‘SEPT9′, or ‘septin 9′, and ‘colorectal cancer’ or ‘colorectal carcinoma’ to identify all relevant studies. 215 studies were identified from MEDLINE and EMBASE, and 151 relevant articles were identified from CMBdisc and CJFD (Fig. 1).
Study inclusion and exclusion criteria
The aim of the study selection is to identify the studies that are clinical studies evaluating the performance of the SEPT9 assay using blood samples from human subjects. (Fig. 1). Duplicates from all databases were excluded and 180 studies remained. In the next screening, a total of 104 articles, including letters, reviews, meta-analysis and guidelines (18 in MEDLINE and EMBASE, 61 in CBMdisc and CJFD), basic research studies (3 in MEDLINE and EMBASE, 7 in CBMdisc and CJFD) and articles irrelevant to mSEPT9 detection assays (3 in MEDLINE and EMBASE, 12 in CBMdisc and CJFD) were excluded. In the following eligibility screening, a further 48 articles were excluded, including studies that were not using plasma or serum samples (16 in MEDLINE and EMBASE, 5 in CBMdisc and CJFD), studies that were not detecting gene methylation (9 in MEDLINE and EMBASE, 18 in CBMdisc and CJFD). Finally, 28 studies were included in the qualitative synthesis, and studies that did not have statistically significant number of CRC or non-CRC cases included (3 in CBMdisc and CJFD), and 25 studies were included in the quantitative synthesis for this meta-analysis.
Data extraction
A standardized data abstraction form was developed, and key elements related to test parameters were collected by two independent reviewers. The study design, type of study (case-control, cohort or screening), sample types, sample size (number of cases, controls, males, females), subject age distribution, algorithm and gold standard for diagnosis were first examined and recorded to ensure the validity of a study. The test sensitivity, specificity and positivity rate with detailed positive, negative and total numbers of cases were then collected. The PLR, NLR and OR were calculated based on these numbers.
Study quality and risk of bias assessment
The quality of the included reports was assessed using the QUADAS system8 by Review Manager 5.2 software. The methodological quality of the studies with focuses on the risk of bias and applicability was systematically assessed, as shown in Supplementary Figure 1. All items in the PRISMA checklist was assessed and finished, as shown in Supplementary Table 1. Most studies were cohort or case-control studies, and only three of them were prospective screening studies. Therefore, some bias inevitably appeared in cohort or case-control studies. Although most of them clearly describe the patient selection criteria and procedure, and the number of CRC cases was comparable with the number of controls, a few of them were lack of description of selection criteria, or had unbalanced number of cases against controls, and these may lead to the risk of bias. As the kits from Epigenomics, Inc. were used for a majority of studies, the cutoff values were identical for most studies, and the index test (marker test) and the reference test were performed parallel. Finally, colonoscopy and subsequent pathological examination were used as the gold standard for cancer or normal subject determination for all studies, and this ensured the quality of data. The overall bias of the included studies was tested using the Deeks’ funnel plot (Fig. 6), and the P value of 0.77 indicates that the distribution of studies is symmetric and there is no systematic bias across all studies analyzed in this study.
Subgroup analyses and definition of diagnostic outcomes
All studies were divided into four subgroups based on the algorithm used in the interpretation of multiple qRT-PCR or high-resolution melting (HRM) data. If the final positive test result was determined from at least one positive count out of three repeats, the study was categorized into the 1/3 algorithm group, and if the final positive test result was determined from at least two positive counts out of three repeats, the study was categorized into the 2/3 algorithm group. Similarly, if the final positive test result was determined from at least one positive count out of two repeats, the study was categorized into the 1/2 algorithm group, while if the final positive test result was determined from only one reaction, the study was categorized into the 1/1 algorithm group. The sensitivity, specificity, PLR, NLR and OR were calculated based on the algorithm, and parameters for each algorithm were calculated by pooling the data from studies in the same algorithm group. The stage-dependent sensitivity was calculated based on the number of positive cases and the total number of cases for a certain stage and the algorithm used in the study.
The heterogeneity of the studies was analyzed based on four algorithms used for SEPT9 assay interpretation, and RR (relative risk) and OR (odds ratio) values were calculated to show the heterogeneity in all four algorithms. It can be seen from Fig. 7 that all RR analysis (the left figure in each panel) showed high I2 values (69.3% to 92.6%) with very small P values, indicating the existence of heterogeneity in studies for all four algorithms. The OR analysis (the right figure in each panel) showed high I2 values (54.3–77.0%) with very small P values in 1/3. 2/3 and 1/1 algorithm, suggesting the presence of heterogeneity, while did not show heterogeneity for 1/2 algorithm. Taken together, it can be suggested that heterogeneity was present among studies in all algorithms, and the random effect model should be used for analysis.
Statistical analysis
The sROC curves for each algorithm were simulated, and the Forest plot for each algorithm and the relevant statistics (including estimated sensitivity, specificity, area under curve (AUC), 95% confidence interval, 95% confidence contour, and 95% prediction contour) were performed using the Stata 14.0 software. The Forest plot for the five serum protein markers and relevant statistics were performed using the Meta-disc 1.4 software (Supplementary Figures 2– 6). The estimates and 95% confidence interval for all data in histograms were calculated and plotted using the PRISM 5.0 software. The comparison of ratio parameters was performed using the χ2 test, and p < 0.05 was regarded as a significant difference.
References
National Cancer Institute. Survival rate for colorectal cancer by stage, Physician Data Query, Treatment, Health Professionals 1999 (1999).
Lin, J. S. et al. Screening for Colorectal Cancer: Updated Evidence Report and Systematic Review for the US Preventive Services Task Force. JAMA 315, 2576–94 (2016).
Lofton-Day, C. et al. DNA methylation biomarkers for blood-based colorectal cancer screening. Clin Chem 54, 414–23 (2008).
Grützmann, R. et al. Sensitive detection of colorectal cancer in peripheral blood by septin 9 DNA methylation assay. PLoS One 3, e3759 (2008).
Wasserkort, R. et al. Aberrant septin 9 DNA methylation in colorectal cancer is restricted to a single CpG island. BMC Cancer 13, 398 (2013).
Tóth, K. et al. Detection of methylated SEPT9 in plasma is a reliable screening method for both left- and right-sided colon cancers. PLoS One 7, e46000 (2012).
Church, T. R. et al. Prospective evaluation of methylated SEPT9 in plasma for detection of asymptomatic colorectal cancer. Gut 63, 317–25 (2014).
Whiting, P. et al. The development of QUADAS: a tool for the quality assessment of studies of diagnostic accuracy included in systematic reviews. BMC Med Res Methodol 3, 25 (2003).
Potter, N. T. et al. Validation of a real-time PCR-based qualitative assay for the detection of methylated SEPT9 DNA in human plasma. Clin Chem 60, 1183–91 (2014).
Wu, D. et al. Detection of Colorectal Cancer Using a Simplified SEPT9 Gene Methylation Assay Is a Reliable Method for Opportunistic Screening. J Mol Diagn 18, 535–45 (2016).
Song, L. et al. Algorithm Optimization in Methylation Detection with Multiple RT-qPCR. PLoS One 11, e0163333 (2016).
Ahlquist, D. A. et al. The stool DNA test is more accurate than the plasma septin 9 test in detecting colorectal neoplasia. Clin Gastroenterol Hepatol 10, 272–277 (2012).
Johnson, D. A. et al. Plasma Septin9 versus fecal immunochemical testing for colorectal cancer screening: a prospective multicenter study. PLoS One 9, e98238 (2014).
Weiss, G. & Rosch, T. Potential of a new blood test for colorectal cancer screening—the Septin 9 gene biomarker. Eur Oncol 6, 51–54 (2010).
Liu, Y. et al. Serum methylation levels of TAC1. SEPT9 and EYA4 as diagnostic markers for early colorectal cancers: a pilot study. Biomarkers 18, 399–405 (2013).
He, Q. et al. Development of a multiplex MethyLight assay for the detection of multigene methylation in human colorectal cancer. Cancer Genet Cytogenet 202, 1–10 (2010).
Wang, Z. et al. The application of MS-HRM detection of free methylated SEPT9 in early diagnosis of colorectal cancer. Guangdong Medical Journal 33, 1732–1734 (2012).
Li, S. J. et al. The study of peripheral methylation SEPT9 in screening colorectal cancer. Chin J Gen Surg 124, 1756–1760 (2015).
He, Q. et al. Detection of ALX4 and SEPT9 methylation in human colorectal cancer by Multiplex MethyLight assay. J Sun Yat-Sen Univ (Med Sci) 36, 657–662 (2015).
Tóth, K. et al. Detection of methylated septin 9 in tissue and plasma of colorectal patients with neoplasia and the relationship to the amount of circulating cell-free DNA. PLoS One 9, e115415 (2014).
Kang, Q. et al. Significance of Septin9 gene methylation detection of plasma circulation DNA in colorectal cancer screening. Zhonghua Yi Xue Za Zhi 94, 3839–41 (2014).
He, N. et al. Clinical significance of detecting serum methylated Sept9 gene in diagnosis of colorectal cancer. Chin J Dig 34, 726–731 (2014).
Jin, P. et al. Performance of a second-generation methylated SEPT9 test in detecting colorectal neoplasm. J Gastroenterol Hepatol 30, 830–3 (2015).
Yu, D., Zhang, X. H. & Lu, X. X. Study on diagnostic value of SEPT9 gene methylation in serum for colorectal cancer. Chin J Clin Lab Sci 33, 687–689 (2015).
Ding, Q. Q. et al. Study of methylation SEPT9 for colorectal cancer screening in aged population. Chin J Geriatr 34, 1348–1350 (2015).
deVos, T. et al. Circulating methylated SEPT9 DNA in plasma is a biomarker for colorectal cancer. Clin Chem 5, 1337–46 (2009).
Tänzer, M. et al. Performance of epigenetic markers SEPT9 and ALX4 in plasma for detection of colorectal precancerous lesions. PLoS One 5, e9061 (2010).
Warren, J. D. et al. Septin 9 methylated DNA is a sensitive and specific blood test for colorectal cancer. BMC Med 9, 133 (2010).
Ørntoft, M. B. et al. Danish Study Group on Early Detection of Colorectal Cancer. Performance of the colorectal cancer screening marker Sept9 is influenced by age, diabetes and arthritis: a nested case-control study. BMC Cancer 15, 819 (2015).
Moher, D., Liberati, A., Tetzlaff, J. & Altman, D. G. & The PRISMA Group. Preferred Reporting Items for Systematic Reviews and Meta-Analyses: The PRISMA Statement. PLoS. Med. 6, e1000097 (2009).
Liberati, A. et al. The PRISMA Statement for Reporting Systematic Reviews and Meta-Analyses of Studies That Evaluate Health Care Interventions: Explanation and Elaboration. PLoS. Med. 6, e1000100 (2009).
Liu, Z. et al. A systematic review and meta-analysis of diagnostic and prognostic serum biomarkers of colorectal cancer. PLoS One 9, e103910 (2014).
Lee, J. K. et al. Accuracy of fecal immunochemical tests for colorectal cancer: systematic review and meta-analysis. Ann Intern Med 160, 171 (2014).
Imperiale, T. F. et al. Multitarget stool DNA testing for colorectal-cancer screening. N Engl J Med 370, 1287–97 (2014).
Adler, A. et al. Improving compliance to colorectal cancer screening using blood and stool based tests in patients refusing screening colonoscopy in Germany. BMC Gastroenterol 14, 183 (2014).
Mayo, E., Kinzler, M. & Zhang, L. Considerations for Conducting Meta-analysis in Diagnostic Pathology. Arch Pathol Lab Med 139, 1331 (2015).
Marchevsky, A. M. & Wick, M. R. Evidence-based pathology: systematic literature reviews as the basis for guidelines and best practices. Arch Pathol Lab Med 139, 394–399 (2015).
Acknowledgements
This work was supported by the Beijing Municipal Science and Technology Project (capital public health project) No. Z151100003915092 sponsored by the Beijing Municipal Science and Technology Commission.
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Lele Song and Yuemin Li conceived and designed the study. Lele Song, Jia Jia and Xiumei Peng performed the literature search, collected and analyzed data. Lele Song made tables, figures and wrote the manuscript. Wenhua Xiao and Yuemin Li proof-read the manuscript and made corrections. Lele Song submitted the manuscript.
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Song, L., Jia, J., Peng, X. et al. The performance of the SEPT9 gene methylation assay and a comparison with other CRC screening tests: A meta-analysis. Sci Rep 7, 3032 (2017). https://doi.org/10.1038/s41598-017-03321-8
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DOI: https://doi.org/10.1038/s41598-017-03321-8
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