Introduction 1 State the marker examined， the study objectives， and any prespecified Materials and methodsPatients2 Describe the characteristics ( disease stage or comorbidities) of the study patients， including their source and inclusion and exclusion 3 Describe treatments received and how chosen ( randomized or rule-based) Specimen characteristics4 Describe type of biological material used (including control samples) and methods of preservation and Assay methods5 Specify the assay method used and provide (or reference) a detailed protocol， including specific reagents or kits used， quality control procedures， reproducibility assessments， quantitation methods， and scoring and reporting Specify whether and how assays were performed blinded to the study Study design6 State the method of case selection， including whether prospective or retrospective and whether stratification or matching ( by stage of disease or age) was Specify the time period from which cases were taken， the end of the follow-up period， and the median follow-up 7 Precisely define all clinical endpoints 8 List all candidate variables initially examined or considered for inclusion in 9 Give rationale for sample size; if the study was designed to detect a specified effect size， give the target power and effect Statistical analysis methods10 Specify all statistical methods， including details of any variable selection procedures and other model-building issues， how model assumptions were verified， and how missing data were 11 Clarify how marker values were handled in the analyses; if relevant， describe methods used for cutpoint ResultsData 12 Describe the flow of patients through the study， including the number of patients included in each stage of the analysis (a diagram may be helpful) and reasons for Specifically， both overall and for each subgroup extensively examined report the numbers of patients and the number of 13 Report distributions of basic demographic characteristics (at least age and sex)， standard (disease-specific) prognostic variables， and tumor marker， including numbers of missing Analysis and presentation 14 Show the relation of the marker to standard prognostic 15 Present univariate analyses showing the relation between the marker and outcome， with the estimated effect ( hazard ratio and survival probability) Preferably provide similar analyses for all other variables being For the effect of a tumor marker on a time-to-event outcome， a Kaplan–Meier plot is 16 For key multivariable analyses， report estimated effects ( hazard ratio) with confidence intervals for the marker and， at least for the final model， all other variables in the 17 Among reported results， provide estimated effects with confidence intervals from an analysis in which the marker and standard prognostic variables are included， regardless of their statistical 18 If done， report results of further investigations， such as checking assumptions， sensitivity analyses， and internal Discussion19 Interpret the results in the context of the prespecified hypotheses and other relevant studies; include a discussion of limitations of the 20 Discuss implications for future research and clinical
