From cd0564d20a961648f5bafcb28f4e1e15e5b807b3 Mon Sep 17 00:00:00 2001 From: Lukas Forer Date: Fri, 1 Mar 2024 13:03:42 +0100 Subject: [PATCH 01/10] Add rmd report for a single sample and add versions to Docker --- Dockerfile | 13 ++- modules/local/report.nf | 3 + modules/local/sample_report.nf | 45 ++++++++ reports/report.Rmd | 32 ++++-- reports/sample.Rmd | 192 +++++++++++++++++++++++++++++++++ workflows/mtdna_server_2.nf | 20 +++- 6 files changed, 291 insertions(+), 14 deletions(-) create mode 100644 modules/local/sample_report.nf create mode 100644 reports/sample.Rmd diff --git a/Dockerfile b/Dockerfile index 3b67a71..3439f30 100644 --- a/Dockerfile +++ b/Dockerfile @@ -13,9 +13,10 @@ RUN \ # Install mutserve (not as conda package available) +ENV MUTSERV_VERSION=2.0.0-rc12 RUN mkdir /opt/mutserve WORKDIR "/opt/mutserve" -#RUN wget https://github.com/seppinho/mutserve/releases/download/v2.0.0-rc12/mutserve.zip +#RUN wget https://github.com/seppinho/mutserve/releases/download/v${MUTSERV_VERSION}/mutserve.zip COPY files/mutserve.zip . RUN unzip mutserve.zip && \ rm mutserve.zip @@ -24,18 +25,20 @@ ENV PATH="/opt/mutserve:${PATH}" # Install haplocheck 1.3.3 +ENV HAPLOCHECK_VERSION=1.3.3 RUN mkdir /opt/haplocheck WORKDIR "/opt/haplocheck" -RUN wget https://github.com/genepi/haplocheck/releases/download/v1.3.3/haplocheck.zip +RUN wget https://github.com/genepi/haplocheck/releases/download/v${HAPLOCHECK_VERSION}/haplocheck.zip RUN unzip haplocheck.zip && \ rm haplocheck.zip ENV PATH="/opt/haplocheck:${PATH}" +ENV HAPLOGREP_VERSION=3.2.1 RUN mkdir /opt/haplogrep WORKDIR "/opt/haplogrep" -RUN wget https://github.com/genepi/haplogrep3/releases/download/v3.2.1/haplogrep3-3.2.1-linux.zip && \ - unzip haplogrep3-3.2.1-linux.zip && \ - rm haplogrep3-3.2.1-linux.zip +RUN wget https://github.com/genepi/haplogrep3/releases/download/v${HAPLOGREP_VERSION}/haplogrep3-${HAPLOGREP_VERSION}-linux.zip && \ + unzip haplogrep3-${HAPLOGREP_VERSION}-linux.zip && \ + rm haplogrep3-${HAPLOGREP_VERSION}-linux.zip ENV PATH="/opt/haplogrep:${PATH}" WORKDIR "/opt" diff --git a/modules/local/report.nf b/modules/local/report.nf index 7c0fac6..85529ed 100644 --- a/modules/local/report.nf +++ b/modules/local/report.nf @@ -26,6 +26,9 @@ process REPORT { echo -e "Base Quality\t${params.baseQ}" >> params.txt echo -e "Map Quality\t${params.mapQ}" >> params.txt echo -e "Alignment Quality\t${params.alignQ}" >> params.txt + echo -e "Mutserv\t\${MUTSERV_VERSION}" >> params.txt + echo -e "Haplocheck\t\${HAPLOCHECK_VERSION}" >> params.txt + echo -e "Haplogrep\t\${HAPLOGREP_VERSION}" >> params.txt Rscript -e "require('rmarkdown'); render('${report}', params = list( diff --git a/modules/local/sample_report.nf b/modules/local/sample_report.nf new file mode 100644 index 0000000..c708aeb --- /dev/null +++ b/modules/local/sample_report.nf @@ -0,0 +1,45 @@ +process SAMPLE_REPORT { + + publishDir "${params.output_reports}/samples", mode: 'copy' + + input: + path report + path variants + path haplogroups + path haplocheck + path statistics + path mapping + path excluded + val sample + + output: + file "*.html" + + """ + echo -e "Parameter\tValue" > params.txt + echo -e "Version\t${workflow.manifest.version}" >> params.txt + echo -e "Job\t${params.project}" >> params.txt + echo -e "Date\t${params.project_date}" >> params.txt + echo -e "Repository\t${params.service.github}" >> params.txt + echo -e "Variant Caller\t${params.mode}" >> params.txt + echo -e "Detection Limit\t${params.detection_limit}" >> params.txt + echo -e "Reference\t${params.reference}" >> params.txt + echo -e "Base Quality\t${params.baseQ}" >> params.txt + echo -e "Map Quality\t${params.mapQ}" >> params.txt + echo -e "Alignment Quality\t${params.alignQ}" >> params.txt + + Rscript -e "require('rmarkdown'); render('${report}', + params = list( + pipeline_parameters = 'params.txt', + variants = '${variants}', + haplogroups = '${haplogroups}', + haplocheck = '${haplocheck}', + statistics = '${statistics}', + mapping = '${mapping}', + excluded_samples = '${excluded}', + sample = '${sample}' + ), + knit_root_dir='\$PWD', output_file='\$PWD/${sample}.html')" + """ + +} \ No newline at end of file diff --git a/reports/report.Rmd b/reports/report.Rmd index 349262d..fc0f096 100644 --- a/reports/report.Rmd +++ b/reports/report.Rmd @@ -35,6 +35,10 @@ knitr::opts_chunk$set(echo = FALSE) ```{r echo=FALSE, results='asis'} +link_to_sample <- function(sample) { + paste0('', sample,'' ) +} + variants <- read.delim(params$variants) contamination <- read.delim(params$haplocheck) haplogroups <- read.delim(params$haplogroups) @@ -131,12 +135,15 @@ colors <- c("#28a745", "#dc3545") names(colors) <- c("PASSED", "FAILED") statistics %>% - select("Sample_Label", "MeanDepth", "CoveragePercentage", "CoveredBases", "MeanBaseQuality", "MeanMapQuality", "qc") %>% + mutate (Sample_Link = link_to_sample(Sample_Label)) %>% + select("Sample_Link", "MeanDepth", "CoveragePercentage", "CoveredBases", "MeanBaseQuality", "MeanMapQuality", "qc") %>% datatable( colnames=c("Sample", "Mean Coverage", "Covered Bases (%)", "Covered Bases", "Mean Base Quality", "Mean Mapping Quality", "QC"), options = list( bPaginate = FALSE - ) + ), + escape = FALSE, + class = 'cell-border stripe' ) %>% formatStyle( 'qc', @@ -194,12 +201,15 @@ ggplotly( ```{r echo=FALSE, results='asis'} contaminated_samples %>% - select("Sample_Label", "Contamination_Status", "Contamination.Level") %>% + mutate (Sample_Link = link_to_sample(Sample_Label)) %>% + select("Sample_Link", "Contamination_Status", "Contamination.Level") %>% datatable( colnames=c("Sample", "Contamination","Level"), options = list( bPaginate = FALSE - ) + ), + escape = FALSE, + class = 'cell-border stripe' ) ``` @@ -214,12 +224,15 @@ Row ```{r echo=FALSE, results='asis'} variants_count %>% - select("Sample_Label", "N") %>% + mutate (Sample_Link = link_to_sample(Sample_Label)) %>% + select("Sample_Link", "N") %>% datatable( colnames=c("Sample", "Number of Variants"), options = list( bPaginate = FALSE - ) + ), + escape = FALSE, + class = 'cell-border stripe' ) ``` @@ -271,12 +284,15 @@ Row ```{r echo=FALSE, results='asis'} haplogroups %>% - select("Sample_Label", "Haplogroup", "Quality") %>% + mutate (Sample_Link = link_to_sample(Sample_Label)) %>% + select("Sample_Link", "Haplogroup", "Quality") %>% datatable( colnames=c("Sample", "Haplogroup", "Quality"), options = list( bPaginate = FALSE - ) + ), + escape = FALSE, + class = 'cell-border stripe' ) ``` diff --git a/reports/sample.Rmd b/reports/sample.Rmd new file mode 100644 index 0000000..12f238a --- /dev/null +++ b/reports/sample.Rmd @@ -0,0 +1,192 @@ +--- +title: "`r params$sample`" +output: + html_document: + + +params: + haplogroups: ../tests/data/report/haplogroups.txt + haplocheck: ../tests/data/report/haplocheck.txt + variants: ../tests/data/report/variants_ann.txt + statistics: ../tests/data/report/sample_statistics.txt + mapping: ../tests/data/report/sample_mappings.txt + excluded_samples: ../tests/data/report/excluded_samples.txt + pipeline_parameters: ../tests/data/report/params.txt + sample: HG00180 +--- + +```{css, echo=FALSE} +.level3 { + margin-top: 40px; + border: 1px solid #cccccc; + border-radius: 5px; + padding-bottom: 0px; + margin-bottom: 0px; +} + +.level3 h3 { + margin-top: 0px; + padding-top: 15px; + padding-left: 20px; + padding-right: 20px; + padding-bottom: 15px; + border-bottom: 1px solid #cccccc; + background-color: #00000008; +} +.level3 p { + margin-top: 20px; + padding-left: 20px; + padding-right: 20px; + font-size: 15px; +} +``` + +```{r setup, include=FALSE} + + +MIN_COVERAGE_PERCENTAGE = 90; +MIN_MEAN_BASE_QUALITY = 15; +MIN_MEAN_DEPTH = 50 + +library(ggplot2) +library(DT) +library(tidyverse) +library(data.table) +library(plotly) +library(flexdashboard) +library(knitr) + +knitr::opts_chunk$set(echo = FALSE) +``` + +```{r echo=FALSE, results='asis'} + +link_to_sample <- function(sample) { + paste0('', sample,'' ) +} + +link_to_variant <- function(pos, ref, alt) { + paste0('',pos, ' (', ref, '>', alt,')') +} + +create_label <- function(text, color) { + paste0('', text, '') +} + +variants <- read.delim(params$variants) +contamination <- read.delim(params$haplocheck) +haplogroups <- read.delim(params$haplogroups) +statistics <- read.delim(params$statistics) +pipeline_params <- read.delim(params$pipeline_parameters) + +excluded_samples <- try(read.delim(params$excluded_samples, header = FALSE, )) +if(inherits(excluded_samples, "try-error")) { + excluded_samples = data.frame(V1=c()) +} + + +mapping <- read.delim(params$mapping) +mapping <- mapping %>% + rename(Sample_Label = Sample) +mapping <- mapping %>% + mutate( + qc = case_when( + Filename %in% excluded_samples$V1 ~ "FAILED", + TRUE ~ "PASSED" + ) + ) + + +statistics <- merge(statistics, mapping, by.x="Sample", by.y="Filename") %>% + filter(Sample_Label == params$sample) + + +haplogroups <- merge(haplogroups, mapping, by.x="SampleID", by.y="Filename") %>% + filter(Sample_Label == params$sample) + +contamination <- merge(contamination, mapping, by.x="Sample", by.y="Filename") %>% + filter(Sample_Label == params$sample) + +contaminated_samples = contamination %>% + rename("Contamination_Status" = "Contamination.Status") %>% + filter(Sample_Label == params$sample) + +variants_count <- data.table(t(table(variants$ID))) +variants_count <- merge(variants_count, mapping, by.x="V2", by.y="Filename") %>% + filter(Sample_Label == params$sample) + +variants <- merge(variants, mapping, by.x="ID", by.y="Filename") %>% + filter(Sample_Label == params$sample) + +variants <- variants %>% + mutate(Type = case_when( + Type == "0/1" ~ "2", + Type == "1/0" ~ "2", + Type == "INDEL" ~ "3", + TRUE ~ as.character(Type) + )) %>% + mutate(Type_Label = case_when( + Type == 1 ~ create_label('Variant', 'primary'), + Type == 2 ~ create_label('Heteroplasmy', 'default'), + Type == 3 ~ create_label('InDel', 'info'), + TRUE ~ "Unknown" + )) + +variant_caller <- pipeline_params %>% + filter(Parameter == "Variant Caller") + +``` + + +### Summary + +**Haplogroup:** `r haplogroups$Haplogroup` (`r haplogroups$Quality`) + +**Contaminated:** `r contaminated_samples$Contamination_Status` + + +### Statistics + +```{r echo=FALSE} +statistics %>% + select("Parameter", "Value") %>% + datatable( + options = list( + bPaginate = FALSE, + bFilter = FALSE, + bSort = FALSE, + info = FALSE + ), + escape = FALSE, + rownames = FALSE, + class = 'cell-border stripe' + ) +``` + + + +### Variants and Hetroplasmies + +Detected **`r variants %>% filter(Type_Label == "Variant") %>% nrow()` variants**, **`r variants %>% filter(Type_Label == "Heteroplasmie") %>% nrow()` heteroplasmies** and **`r variants %>% filter(Type_Label == "Indel") %>% nrow()` InDels**. + +```{r echo=FALSE} +variants %>% + mutate( + Variant_Label = link_to_variant(Pos, Ref, Variant) + ) %>% + select("Variant_Label", "VariantLevel", "Type_Label") %>% + datatable( + colnames=c("Variant", "Level", ""), + options = list( + bPaginate = FALSE, + bFilter = FALSE, + bSort = FALSE, + bFooter = FALSE, + info = FALSE + ), + escape = FALSE, + rownames = FALSE, + class = 'cell-border stripe' + ) +``` + diff --git a/workflows/mtdna_server_2.nf b/workflows/mtdna_server_2.nf index b11588c..9fb3bec 100644 --- a/workflows/mtdna_server_2.nf +++ b/workflows/mtdna_server_2.nf @@ -27,11 +27,13 @@ include { VCF_MERGE } from '../modules/local/vcf_merge' include { ANNOTATE } from '../modules/local/annotate' include { HAPLOGROUPS_CONTAMINATION } from '../modules/local/haplogroups_contamination' include { REPORT } from '../modules/local/report' +include { SAMPLE_REPORT } from '../modules/local/sample_report' workflow MTDNA_SERVER_2 { - report_file_ch = file("$projectDir/reports/report.Rmd") + report_file_ch = file("$projectDir/reports/report.Rmd", checkIfExists:true) + sample_report_file_ch = file("$projectDir/reports/sample.Rmd", checkIfExists:true) bams_ch = Channel.fromPath(params.files, checkIfExists:true) if(params.reference.equals("rcrs")){ @@ -172,6 +174,22 @@ workflow MTDNA_SERVER_2 { INPUT_VALIDATION.out.excluded_ch ) + samples = INPUT_VALIDATION.out.mapping_ch + .splitCsv(header: true, sep: '\t') + .map { row -> row.Sample } + + SAMPLE_REPORT( + sample_report_file_ch, + variants_txt_ch, + haplogrep_ch, + contamination_ch, + INPUT_VALIDATION.out.summarized_ch, + INPUT_VALIDATION.out.mapping_ch, + INPUT_VALIDATION.out.excluded_ch, + samples + ) + + } workflow.onComplete { From 0ee0d66542bf4e58470292f644cb226ae9ebace7 Mon Sep 17 00:00:00 2001 From: Lukas Forer Date: Fri, 1 Mar 2024 13:06:07 +0100 Subject: [PATCH 02/10] Fix number of tasks in snapshot --- tests/mitocalling.nf.test.snap | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/tests/mitocalling.nf.test.snap b/tests/mitocalling.nf.test.snap index 6d9e99b..a6609fd 100644 --- a/tests/mitocalling.nf.test.snap +++ b/tests/mitocalling.nf.test.snap @@ -22,8 +22,8 @@ "variants.annotated.txt:md5,3946a3bd26ceafe8f61903499bfcbb35", { "tasksFailed": 0, - "tasksCount": 10, - "tasksSucceeded": 10 + "tasksCount": 12, + "tasksSucceeded": 12 } ], "timestamp": "2024-02-28T12:32:40.624776" From b375e7d2f93ee7f6c9b8f6c569427ff2a78df27f Mon Sep 17 00:00:00 2001 From: Lukas Forer Date: Fri, 1 Mar 2024 14:47:24 +0100 Subject: [PATCH 03/10] Add testcase for 6 samples --- tests/lib/DownloadUtil.groovy | 97 ++++++++++++++++++++++++++++++++++ tests/mitocalling.nf.test | 28 +++++++++- tests/mitocalling.nf.test.snap | 31 +++++++++++ 3 files changed, 154 insertions(+), 2 deletions(-) create mode 100644 tests/lib/DownloadUtil.groovy diff --git a/tests/lib/DownloadUtil.groovy b/tests/lib/DownloadUtil.groovy new file mode 100644 index 0000000..b5860b8 --- /dev/null +++ b/tests/lib/DownloadUtil.groovy @@ -0,0 +1,97 @@ + +@Grab(group='org.apache.httpcomponents', module='httpclient', version='4.5.13') +@Grab(group='org.apache.httpcomponents', module='httpmime', version='4.5.13') + +import org.apache.http.client.methods.HttpGet +import org.apache.http.impl.client.HttpClients +import org.apache.http.client.config.RequestConfig +import org.apache.http.client.utils.URIBuilder +import org.apache.http.entity.ContentType +import org.apache.http.HttpEntity +import org.apache.http.util.EntityUtils + +import java.nio.file.Files +import java.nio.file.Paths +import java.nio.file.StandardCopyOption +import java.util.zip.ZipInputStream +import java.io.FileOutputStream + +public class DownloadUtil { + + public static void downloadAndExtractZip(String url, String destinationFolder) { + // Create HTTP client + def httpClient = HttpClients.custom().setDefaultRequestConfig(RequestConfig.custom().setConnectTimeout(5000).build()).build() + + try { + // Prepare HTTP GET request + def uri = new URIBuilder(url).build() + def httpGet = new HttpGet(uri) + + // Execute HTTP request + def response = httpClient.execute(httpGet) + + // Check if response is successful + if (response.getStatusLine().getStatusCode() == 200) { + // Get HTTP entity + def entity = response.getEntity() + + // Create temporary file to save the downloaded zip + def tempFile = Files.createTempFile("temp-", ".zip") + + // Save the zip file + Files.copy(entity.getContent(), tempFile, StandardCopyOption.REPLACE_EXISTING) + + // Extract the zip file + extractZip(tempFile.toString(), destinationFolder) + + } else { + println "Failed to download the zip file. HTTP status code: ${response.getStatusLine().getStatusCode()}" + } + } finally { + // Close HTTP client + httpClient.close() + } + } + + public static void extractZip(String zipFilePath, String destinationFolder) { + try { + // Open zip input stream + def zipInputStream = new ZipInputStream(new FileInputStream(zipFilePath)) + + // Create destination folder if it does not exist + Files.createDirectories(Paths.get(destinationFolder)) + + // Extract each entry in the zip file + def entry = zipInputStream.getNextEntry() + while (entry != null) { + def entryName = entry.getName() + def outputFile = Paths.get(destinationFolder, entryName) + + // If entry is a directory, create it + if (entry.isDirectory()) { + Files.createDirectories(outputFile) + } else { + // Create parent directory if it does not exist + Files.createDirectories(outputFile.getParent()) + + // Extract file + def outputStream = new FileOutputStream(outputFile.toFile()) + byte[] buffer = new byte[4096] + def len + while ((len = zipInputStream.read(buffer)) > 0) { + outputStream.write(buffer, 0, len) + } + outputStream.close() + } + + entry = zipInputStream.getNextEntry() + } + + zipInputStream.closeEntry() + zipInputStream.close() + } catch (Exception e) { + println "Error extracting zip file: ${e.message}" + } + } + +} \ No newline at end of file diff --git a/tests/mitocalling.nf.test b/tests/mitocalling.nf.test index 6a55f5d..4b68821 100644 --- a/tests/mitocalling.nf.test +++ b/tests/mitocalling.nf.test @@ -42,7 +42,7 @@ nextflow_pipeline { } - test("Runs with single BAM file and mutect2") { + test("Runs with single BAM file and mutect2") { when { params { @@ -63,7 +63,7 @@ nextflow_pipeline { } - test("Runs with two files and one goes through QC") { + test("Runs with two files and one goes through QC") { when { params { @@ -83,4 +83,28 @@ nextflow_pipeline { } + test("Runs with 6 samples provided as testdata") { + + setup { + DownloadUtil.downloadAndExtractZip("https://mitoverse.i-med.ac.at/downloads/6-samples.zip", "${launchDir}/input") + } + + when { + params { + project = "test-job" + files = "${launchDir}/input/*.bam" + mode = "mutect2" + output = "out" + output_reports = "reports" + output_auxiliary = "files" + } + } + + then { + assert workflow.success + assert snapshot(path("${launchDir}/out/variants.annotated.txt"),workflow.trace).match() + } + + } + } \ No newline at end of file diff --git a/tests/mitocalling.nf.test.snap b/tests/mitocalling.nf.test.snap index a6609fd..96415dc 100644 --- a/tests/mitocalling.nf.test.snap +++ b/tests/mitocalling.nf.test.snap @@ -3,20 +3,47 @@ "content": [ "variants.annotated.txt:md5,94fa4b4d0613f61410618a7eba4c5c97" ], + "meta": { + "nf-test": "0.8.4", + "nextflow": "23.04.3" + }, "timestamp": "2024-02-26T23:09:58.251860914" }, "Runs with single BAM file and mutserve": { "content": [ "variants.annotated.txt:md5,aac0aee87ce1a91f8c35264be248b52e" ], + "meta": { + "nf-test": "0.8.4", + "nextflow": "23.04.3" + }, "timestamp": "2024-02-27T15:19:15.934593072" }, "Runs with BAM file including different header contigs": { "content": [ "variants.annotated.txt:md5,d68e6fed173c766228164438a24ecf5a" ], + "meta": { + "nf-test": "0.8.4", + "nextflow": "23.04.3" + }, "timestamp": "2024-02-26T23:03:07.470112898" }, + "Runs with 6 samples provided as testdata": { + "content": [ + "variants.annotated.txt:md5,efeb591e666bd107fbb0ce47898b6304", + { + "tasksFailed": 0, + "tasksCount": 24, + "tasksSucceeded": 24 + } + ], + "meta": { + "nf-test": "0.8.4", + "nextflow": "23.04.3" + }, + "timestamp": "2024-03-01T13:54:57.136897" + }, "Runs with two files and one goes through QC": { "content": [ "variants.annotated.txt:md5,3946a3bd26ceafe8f61903499bfcbb35", @@ -26,6 +53,10 @@ "tasksSucceeded": 12 } ], + "meta": { + "nf-test": "0.8.4", + "nextflow": "23.04.3" + }, "timestamp": "2024-02-28T12:32:40.624776" } } \ No newline at end of file From 79ae1e9b1c5e6ed5b1b1abff317dc13a85a30dd6 Mon Sep 17 00:00:00 2001 From: Sebastian Schoenherr Date: Fri, 1 Mar 2024 16:22:36 +0100 Subject: [PATCH 04/10] Test different modi --- tests/mitocalling.nf.test | 50 ++++++++++++++++++++++++++++++++++++++- 1 file changed, 49 insertions(+), 1 deletion(-) diff --git a/tests/mitocalling.nf.test b/tests/mitocalling.nf.test index 4b68821..7c36514 100644 --- a/tests/mitocalling.nf.test +++ b/tests/mitocalling.nf.test @@ -83,7 +83,31 @@ nextflow_pipeline { } - test("Runs with 6 samples provided as testdata") { + test("Runs with 6 samples provided as testdata with mutserve") { + + setup { + DownloadUtil.downloadAndExtractZip("https://mitoverse.i-med.ac.at/downloads/6-samples.zip", "${launchDir}/input") + } + + when { + params { + project = "test-job" + files = "${launchDir}/input/*.bam" + mode = "mutserve" + output = "out" + output_reports = "reports" + output_auxiliary = "files" + } + } + + then { + assert workflow.success + assert snapshot(path("${launchDir}/out/variants.annotated.txt"),workflow.trace).match() + } + + } + + test("Runs with 6 samples provided as testdata with mutect2") { setup { DownloadUtil.downloadAndExtractZip("https://mitoverse.i-med.ac.at/downloads/6-samples.zip", "${launchDir}/input") @@ -107,4 +131,28 @@ nextflow_pipeline { } + test("Runs with 6 samples provided as testdata with fusion") { + + setup { + DownloadUtil.downloadAndExtractZip("https://mitoverse.i-med.ac.at/downloads/6-samples.zip", "${launchDir}/input") + } + + when { + params { + project = "test-job" + files = "${launchDir}/input/*.bam" + mode = "fusion" + output = "out" + output_reports = "reports" + output_auxiliary = "files" + } + } + + then { + assert workflow.success + assert snapshot(path("${launchDir}/out/variants.annotated.txt"),workflow.trace).match() + } + + } + } \ No newline at end of file From 5c1b9ee3d35bf5ca320686b418d21504d3e7d164 Mon Sep 17 00:00:00 2001 From: Lukas Forer Date: Sat, 2 Mar 2024 10:30:32 +0100 Subject: [PATCH 05/10] Fix warning with empty exclude files --- reports/report.Rmd | 24 ++++++++++++------------ reports/sample.Rmd | 6 +++--- 2 files changed, 15 insertions(+), 15 deletions(-) diff --git a/reports/report.Rmd b/reports/report.Rmd index fc0f096..642ccda 100644 --- a/reports/report.Rmd +++ b/reports/report.Rmd @@ -33,10 +33,10 @@ library(knitr) knitr::opts_chunk$set(echo = FALSE) ``` -```{r echo=FALSE, results='asis'} +```{r echo=FALSE, warning=FALSE, error=FALSE, include=FALSE} link_to_sample <- function(sample) { - paste0('', sample,'' ) + paste0('', sample,'' ) } variants <- read.delim(params$variants) @@ -45,7 +45,7 @@ haplogroups <- read.delim(params$haplogroups) statistics <- read.delim(params$statistics) pipeline_params <- read.delim(params$pipeline_parameters) -excluded_samples <- try(read.delim(params$excluded_samples, header = FALSE, )) +excluded_samples <- try(read.delim(params$excluded_samples, header = FALSE)) if(inherits(excluded_samples, "try-error")) { excluded_samples = data.frame(V1=c()) } @@ -129,7 +129,7 @@ Row ### Summary {data-width=750} -```{r echo=FALSE, results='asis'} +```{r} colors <- c("#28a745", "#dc3545") names(colors) <- c("PASSED", "FAILED") @@ -155,7 +155,7 @@ statistics %>% ### Workflow parameters and Software versions {data-width=250} -```{r echo=FALSE, results='asis'} +```{r} pipeline_params %>% kable() ``` @@ -166,7 +166,7 @@ Row ### Mean Coverage per sample -```{r echo=FALSE, results='asis'} +```{r} ggplotly( ggplot(statistics) + @@ -182,7 +182,7 @@ ggplotly( ### Mean Base Quality per sample -```{r echo=FALSE, results='asis'} +```{r} ggplotly( ggplot(statistics) + geom_col(aes(x=Sample_Label, y=MeanBaseQuality, fill=qc)) + @@ -199,7 +199,7 @@ ggplotly( # Contamination -```{r echo=FALSE, results='asis'} +```{r} contaminated_samples %>% mutate (Sample_Link = link_to_sample(Sample_Label)) %>% select("Sample_Link", "Contamination_Status", "Contamination.Level") %>% @@ -222,7 +222,7 @@ Row ### Summary -```{r echo=FALSE, results='asis'} +```{r} variants_count %>% mutate (Sample_Link = link_to_sample(Sample_Label)) %>% select("Sample_Link", "N") %>% @@ -242,7 +242,7 @@ Row ### Common Variants and Hetereoplasmies -```{r echo=FALSE, results='asis'} +```{r} variants <- variants %>% mutate(Type = case_when( Type == "0/1" ~ "2", @@ -282,7 +282,7 @@ Row ### Summary -```{r echo=FALSE, results='asis'} +```{r} haplogroups %>% mutate (Sample_Link = link_to_sample(Sample_Label)) %>% select("Sample_Link", "Haplogroup", "Quality") %>% @@ -301,7 +301,7 @@ Row ### Quality per Sample -```{r echo=FALSE, results='asis'} +```{r} ggplotly( ggplot(haplogroups) + geom_col(aes(y=Quality, x=Sample_Label), alpha=0.7) + diff --git a/reports/sample.Rmd b/reports/sample.Rmd index 12f238a..3e1b0bc 100644 --- a/reports/sample.Rmd +++ b/reports/sample.Rmd @@ -59,7 +59,7 @@ library(knitr) knitr::opts_chunk$set(echo = FALSE) ``` -```{r echo=FALSE, results='asis'} +```{r echo=FALSE, warning=FALSE, error=FALSE, include=FALSE} link_to_sample <- function(sample) { paste0('', sample,'' ) @@ -147,7 +147,7 @@ variant_caller <- pipeline_params %>% ### Statistics -```{r echo=FALSE} +```{r} statistics %>% select("Parameter", "Value") %>% datatable( @@ -169,7 +169,7 @@ statistics %>% Detected **`r variants %>% filter(Type_Label == "Variant") %>% nrow()` variants**, **`r variants %>% filter(Type_Label == "Heteroplasmie") %>% nrow()` heteroplasmies** and **`r variants %>% filter(Type_Label == "Indel") %>% nrow()` InDels**. -```{r echo=FALSE} +```{r} variants %>% mutate( Variant_Label = link_to_variant(Pos, Ref, Variant) From eb4454674dc8a3ffe27f2ed756d985c6f3b9a956 Mon Sep 17 00:00:00 2001 From: Lukas Forer Date: Sat, 2 Mar 2024 11:02:05 +0100 Subject: [PATCH 06/10] Add contamination status to summary table and commit snapshots --- reports/report.Rmd | 21 ++++++++++++-- tests/mitocalling.nf.test.snap | 50 +++++++++++++++++++++++++++------- 2 files changed, 58 insertions(+), 13 deletions(-) diff --git a/reports/report.Rmd b/reports/report.Rmd index 642ccda..0164a8c 100644 --- a/reports/report.Rmd +++ b/reports/report.Rmd @@ -134,11 +134,21 @@ Row colors <- c("#28a745", "#dc3545") names(colors) <- c("PASSED", "FAILED") +colors_contamination <- c("#dddddd","#28a745", "#dc3545") +names(colors_contamination) <- c("-", "NO", "YES") + statistics %>% - mutate (Sample_Link = link_to_sample(Sample_Label)) %>% - select("Sample_Link", "MeanDepth", "CoveragePercentage", "CoveredBases", "MeanBaseQuality", "MeanMapQuality", "qc") %>% + mutate ( + Sample_Link = link_to_sample(Sample_Label), + contamination = case_when( + qc == "FAILED" ~ "-", + Sample_Label %in% contaminated_samples$Sample_Label ~ "YES", + TRUE ~ "NO" + ) + ) %>% + select("Sample_Link", "MeanDepth", "CoveragePercentage", "CoveredBases", "MeanBaseQuality", "MeanMapQuality", "qc", "contamination") %>% datatable( - colnames=c("Sample", "Mean Coverage", "Covered Bases (%)", "Covered Bases", "Mean Base Quality", "Mean Mapping Quality", "QC"), + colnames=c("Sample", "Mean Coverage", "Covered Bases (%)", "Covered Bases", "Mean Base Quality", "Mean Mapping Quality", "QC", "Cont."), options = list( bPaginate = FALSE ), @@ -149,6 +159,11 @@ statistics %>% 'qc', backgroundColor = styleEqual(c("PASSED", "FAILED"), colors), color = 'white' + ) %>% + formatStyle( + 'contamination', + backgroundColor = styleEqual(c("-", "NO", "YES"), colors_contamination), + color = 'white' ) ``` diff --git a/tests/mitocalling.nf.test.snap b/tests/mitocalling.nf.test.snap index 96415dc..9198b0d 100644 --- a/tests/mitocalling.nf.test.snap +++ b/tests/mitocalling.nf.test.snap @@ -9,6 +9,21 @@ }, "timestamp": "2024-02-26T23:09:58.251860914" }, + "Runs with 6 samples provided as testdata with mutect2": { + "content": [ + "variants.annotated.txt:md5,efeb591e666bd107fbb0ce47898b6304", + { + "tasksFailed": 0, + "tasksCount": 24, + "tasksSucceeded": 24 + } + ], + "meta": { + "nf-test": "0.8.4", + "nextflow": "23.04.3" + }, + "timestamp": "2024-03-02T10:54:19.802653" + }, "Runs with single BAM file and mutserve": { "content": [ "variants.annotated.txt:md5,aac0aee87ce1a91f8c35264be248b52e" @@ -29,34 +44,49 @@ }, "timestamp": "2024-02-26T23:03:07.470112898" }, - "Runs with 6 samples provided as testdata": { + "Runs with two files and one goes through QC": { "content": [ - "variants.annotated.txt:md5,efeb591e666bd107fbb0ce47898b6304", + "variants.annotated.txt:md5,3946a3bd26ceafe8f61903499bfcbb35", { "tasksFailed": 0, - "tasksCount": 24, - "tasksSucceeded": 24 + "tasksCount": 12, + "tasksSucceeded": 12 } ], "meta": { "nf-test": "0.8.4", "nextflow": "23.04.3" }, - "timestamp": "2024-03-01T13:54:57.136897" + "timestamp": "2024-02-28T12:32:40.624776" }, - "Runs with two files and one goes through QC": { + "Runs with 6 samples provided as testdata with fusion": { "content": [ - "variants.annotated.txt:md5,3946a3bd26ceafe8f61903499bfcbb35", + "variants.annotated.txt:md5,b8a0bbf624f5b0ecaac01ca441b50a76", { "tasksFailed": 0, - "tasksCount": 12, - "tasksSucceeded": 12 + "tasksCount": 40, + "tasksSucceeded": 40 } ], "meta": { "nf-test": "0.8.4", "nextflow": "23.04.3" }, - "timestamp": "2024-02-28T12:32:40.624776" + "timestamp": "2024-03-02T10:58:52.06124" + }, + "Runs with 6 samples provided as testdata with mutserve": { + "content": [ + "variants.annotated.txt:md5,5c726eaa61eacfb09c64ebaeb9a2a75a", + { + "tasksFailed": 0, + "tasksCount": 25, + "tasksSucceeded": 25 + } + ], + "meta": { + "nf-test": "0.8.4", + "nextflow": "23.04.3" + }, + "timestamp": "2024-03-02T10:50:44.742545" } } \ No newline at end of file From 9a5d180e43a5c157408d0342771eaba3d311289c Mon Sep 17 00:00:00 2001 From: Lukas Forer Date: Sat, 2 Mar 2024 11:19:45 +0100 Subject: [PATCH 07/10] Fix variant counting and add qc status --- reports/sample.Rmd | 13 ++++++++----- 1 file changed, 8 insertions(+), 5 deletions(-) diff --git a/reports/sample.Rmd b/reports/sample.Rmd index 3e1b0bc..b46e077 100644 --- a/reports/sample.Rmd +++ b/reports/sample.Rmd @@ -12,6 +12,7 @@ params: mapping: ../tests/data/report/sample_mappings.txt excluded_samples: ../tests/data/report/excluded_samples.txt pipeline_parameters: ../tests/data/report/params.txt + #sample: HG00245 sample: HG00180 --- @@ -94,8 +95,8 @@ mapping <- mapping %>% Filename %in% excluded_samples$V1 ~ "FAILED", TRUE ~ "PASSED" ) - ) - + ) %>% + filter(Sample_Label == params$sample) statistics <- merge(statistics, mapping, by.x="Sample", by.y="Filename") %>% filter(Sample_Label == params$sample) @@ -140,9 +141,11 @@ variant_caller <- pipeline_params %>% ### Summary -**Haplogroup:** `r haplogroups$Haplogroup` (`r haplogroups$Quality`) +**Quality Control: ** `r if (mapping$qc == "PASSED") create_label(mapping$qc, 'success') else create_label(mapping$qc, 'danger')` + +**Haplogroup:** `r if (mapping$qc == "PASSED") haplogroups$Haplogroup else '-'` `r if (mapping$qc == "PASSED") paste('(', haplogroups$Quality, ')') else ''` -**Contaminated:** `r contaminated_samples$Contamination_Status` +**Contaminated:** `r if (mapping$qc == "PASSED") contaminated_samples$Contamination_Status else '-'` ### Statistics @@ -167,7 +170,7 @@ statistics %>% ### Variants and Hetroplasmies -Detected **`r variants %>% filter(Type_Label == "Variant") %>% nrow()` variants**, **`r variants %>% filter(Type_Label == "Heteroplasmie") %>% nrow()` heteroplasmies** and **`r variants %>% filter(Type_Label == "Indel") %>% nrow()` InDels**. +Detected **`r variants %>% filter(Type == 1) %>% nrow()` variants**, **`r variants %>% filter(Type == 2) %>% nrow()` heteroplasmies** and **`r variants %>% filter(Type == 3) %>% nrow()` InDels**. ```{r} variants %>% From 34030530ed10862ddeb103623b3a0bae5480b146 Mon Sep 17 00:00:00 2001 From: Lukas Forer Date: Sat, 2 Mar 2024 13:06:26 +0100 Subject: [PATCH 08/10] Add coverage plot to sample report --- modules/local/sample_report.nf | 9 +- reports/sample.Rmd | 15 +- tests/data/report/coverage.tsv | 16579 +++++++++++++++++++++++++++++++ workflows/mtdna_server_2.nf | 7 +- 4 files changed, 16606 insertions(+), 4 deletions(-) create mode 100644 tests/data/report/coverage.tsv diff --git a/modules/local/sample_report.nf b/modules/local/sample_report.nf index c708aeb..b81e35e 100644 --- a/modules/local/sample_report.nf +++ b/modules/local/sample_report.nf @@ -10,7 +10,7 @@ process SAMPLE_REPORT { path statistics path mapping path excluded - val sample + tuple val(sample_filename), val(sample), file(sample_bam_file) output: file "*.html" @@ -28,6 +28,10 @@ process SAMPLE_REPORT { echo -e "Map Quality\t${params.mapQ}" >> params.txt echo -e "Alignment Quality\t${params.alignQ}" >> params.txt + #TODO: split fwd and reverse? https://bioinformatics.stackexchange.com/questions/8649/how-can-i-calculate-coverage-at-single-bases-using-a-bam-file + echo -e "Contig\tPosition\tCoverage" > coverage.tsv + samtools depth -a ${sample_bam_file} >> coverage.tsv + Rscript -e "require('rmarkdown'); render('${report}', params = list( pipeline_parameters = 'params.txt', @@ -37,7 +41,8 @@ process SAMPLE_REPORT { statistics = '${statistics}', mapping = '${mapping}', excluded_samples = '${excluded}', - sample = '${sample}' + sample = '${sample}', + coverage = 'coverage.tsv' ), knit_root_dir='\$PWD', output_file='\$PWD/${sample}.html')" """ diff --git a/reports/sample.Rmd b/reports/sample.Rmd index b46e077..482a734 100644 --- a/reports/sample.Rmd +++ b/reports/sample.Rmd @@ -14,6 +14,7 @@ params: pipeline_parameters: ../tests/data/report/params.txt #sample: HG00245 sample: HG00180 + coverage: ../tests/data/report/coverage.tsv --- ```{css, echo=FALSE} @@ -79,6 +80,7 @@ contamination <- read.delim(params$haplocheck) haplogroups <- read.delim(params$haplogroups) statistics <- read.delim(params$statistics) pipeline_params <- read.delim(params$pipeline_parameters) +coverage <- read.delim(params$coverage) excluded_samples <- try(read.delim(params$excluded_samples, header = FALSE, )) if(inherits(excluded_samples, "try-error")) { @@ -167,6 +169,18 @@ statistics %>% ``` +### Coverage + +```{r, fig.width=9, warning=FALSE} +#ggplotly( + ggplot(coverage) + + geom_hline(yintercept=mean(coverage$Coverage), color="black", linetype="dashed") + + geom_hline(yintercept=mean(coverage$Coverage) + sd(coverage$Coverage), color="gray", linetype="dashed") + + geom_hline(yintercept=mean(coverage$Coverage) - sd(coverage$Coverage), color="gray", linetype="dashed") + + geom_line(aes(x=Position, y=Coverage), color = "darkblue") + + theme_bw() +#) +``` ### Variants and Hetroplasmies @@ -192,4 +206,3 @@ variants %>% class = 'cell-border stripe' ) ``` - diff --git a/tests/data/report/coverage.tsv b/tests/data/report/coverage.tsv new file mode 100644 index 0000000..36d84a7 --- /dev/null +++ b/tests/data/report/coverage.tsv @@ -0,0 +1,16579 @@ +Contig Position Coverage +MT 1 558 +MT 2 565 +MT 3 577 +MT 4 585 +MT 5 600 +MT 6 611 +MT 7 621 +MT 8 630 +MT 9 639 +MT 10 650 +MT 11 657 +MT 12 671 +MT 13 675 +MT 14 683 +MT 15 694 +MT 16 703 +MT 17 713 +MT 18 716 +MT 19 721 +MT 20 729 +MT 21 726 +MT 22 725 +MT 23 730 +MT 24 733 +MT 25 734 +MT 26 737 +MT 27 737 +MT 28 745 +MT 29 744 +MT 30 753 +MT 31 757 +MT 32 770 +MT 33 782 +MT 34 791 +MT 35 793 +MT 36 799 +MT 37 804 +MT 38 808 +MT 39 808 +MT 40 818 +MT 41 829 +MT 42 832 +MT 43 840 +MT 44 854 +MT 45 856 +MT 46 862 +MT 47 877 +MT 48 890 +MT 49 906 +MT 50 916 +MT 51 928 +MT 52 938 +MT 53 948 +MT 54 950 +MT 55 958 +MT 56 961 +MT 57 962 +MT 58 966 +MT 59 975 +MT 60 979 +MT 61 986 +MT 62 993 +MT 63 996 +MT 64 994 +MT 65 1019 +MT 66 1035 +MT 67 1044 +MT 68 1047 +MT 69 1066 +MT 70 1073 +MT 71 1077 +MT 72 1084 +MT 73 1090 +MT 74 1094 +MT 75 1099 +MT 76 1107 +MT 77 1110 +MT 78 1115 +MT 79 1124 +MT 80 1127 +MT 81 1133 +MT 82 1135 +MT 83 1146 +MT 84 1145 +MT 85 1151 +MT 86 1153 +MT 87 1169 +MT 88 1171 +MT 89 1183 +MT 90 1191 +MT 91 1213 +MT 92 1251 +MT 93 1270 +MT 94 1274 +MT 95 1287 +MT 96 1305 +MT 97 1328 +MT 98 1340 +MT 99 1363 +MT 100 1372 +MT 101 1379 +MT 102 1389 +MT 103 1388 +MT 104 1393 +MT 105 1396 +MT 106 1394 +MT 107 1396 +MT 108 1406 +MT 109 1398 +MT 110 1402 +MT 111 1411 +MT 112 1405 +MT 113 1416 +MT 114 1427 +MT 115 1430 +MT 116 1431 +MT 117 1441 +MT 118 1438 +MT 119 1446 +MT 120 1441 +MT 121 1437 +MT 122 1445 +MT 123 1457 +MT 124 1455 +MT 125 1463 +MT 126 1467 +MT 127 1474 +MT 128 1477 +MT 129 1491 +MT 130 1504 +MT 131 1507 +MT 132 1516 +MT 133 1523 +MT 134 1514 +MT 135 1514 +MT 136 1510 +MT 137 1511 +MT 138 1514 +MT 139 1519 +MT 140 1515 +MT 141 1519 +MT 142 1524 +MT 143 1517 +MT 144 1520 +MT 145 1525 +MT 146 1529 +MT 147 1517 +MT 148 1509 +MT 149 1498 +MT 150 1493 +MT 151 1491 +MT 152 1481 +MT 153 1478 +MT 154 1469 +MT 155 1472 +MT 156 1482 +MT 157 1482 +MT 158 1481 +MT 159 1479 +MT 160 1485 +MT 161 1472 +MT 162 1464 +MT 163 1465 +MT 164 1460 +MT 165 1452 +MT 166 1450 +MT 167 1456 +MT 168 1454 +MT 169 1423 +MT 170 1417 +MT 171 1427 +MT 172 1417 +MT 173 1415 +MT 174 1413 +MT 175 1424 +MT 176 1427 +MT 177 1425 +MT 178 1424 +MT 179 1422 +MT 180 1428 +MT 181 1426 +MT 182 1436 +MT 183 1434 +MT 184 1433 +MT 185 1430 +MT 186 1431 +MT 187 1417 +MT 188 1409 +MT 189 1402 +MT 190 1404 +MT 191 1400 +MT 192 1363 +MT 193 1366 +MT 194 1363 +MT 195 1358 +MT 196 1354 +MT 197 1345 +MT 198 1345 +MT 199 1352 +MT 200 1357 +MT 201 1366 +MT 202 1365 +MT 203 1360 +MT 204 1368 +MT 205 1369 +MT 206 1369 +MT 207 1367 +MT 208 1359 +MT 209 1363 +MT 210 1362 +MT 211 1359 +MT 212 1354 +MT 213 1352 +MT 214 1346 +MT 215 1334 +MT 216 1330 +MT 217 1329 +MT 218 1343 +MT 219 1336 +MT 220 1331 +MT 221 1323 +MT 222 1317 +MT 223 1302 +MT 224 1304 +MT 225 1292 +MT 226 1287 +MT 227 1289 +MT 228 1280 +MT 229 1267 +MT 230 1256 +MT 231 1258 +MT 232 1247 +MT 233 1241 +MT 234 1239 +MT 235 1235 +MT 236 1237 +MT 237 1242 +MT 238 1247 +MT 239 1239 +MT 240 1241 +MT 241 1234 +MT 242 1231 +MT 243 1233 +MT 244 1221 +MT 245 1219 +MT 246 1206 +MT 247 1196 +MT 248 1199 +MT 249 1199 +MT 250 1196 +MT 251 1188 +MT 252 1180 +MT 253 1172 +MT 254 1176 +MT 255 1174 +MT 256 1166 +MT 257 1180 +MT 258 1184 +MT 259 1186 +MT 260 1182 +MT 261 1193 +MT 262 1200 +MT 263 1197 +MT 264 1198 +MT 265 1195 +MT 266 1190 +MT 267 1191 +MT 268 1189 +MT 269 1188 +MT 270 1183 +MT 271 1175 +MT 272 1172 +MT 273 1173 +MT 274 1171 +MT 275 1167 +MT 276 1159 +MT 277 1158 +MT 278 1161 +MT 279 1166 +MT 280 1157 +MT 281 1149 +MT 282 1137 +MT 283 1130 +MT 284 1134 +MT 285 1150 +MT 286 1152 +MT 287 1151 +MT 288 1152 +MT 289 1144 +MT 290 1137 +MT 291 1131 +MT 292 1120 +MT 293 1113 +MT 294 1118 +MT 295 1121 +MT 296 1118 +MT 297 1113 +MT 298 1114 +MT 299 1098 +MT 300 1089 +MT 301 1060 +MT 302 1048 +MT 303 1044 +MT 304 1031 +MT 305 1022 +MT 306 1020 +MT 307 1021 +MT 308 1013 +MT 309 997 +MT 310 891 +MT 311 910 +MT 312 853 +MT 313 854 +MT 314 842 +MT 315 869 +MT 316 776 +MT 317 764 +MT 318 742 +MT 319 740 +MT 320 740 +MT 321 747 +MT 322 744 +MT 323 737 +MT 324 743 +MT 325 747 +MT 326 747 +MT 327 757 +MT 328 764 +MT 329 761 +MT 330 760 +MT 331 771 +MT 332 771 +MT 333 779 +MT 334 781 +MT 335 786 +MT 336 784 +MT 337 787 +MT 338 794 +MT 339 797 +MT 340 812 +MT 341 815 +MT 342 825 +MT 343 829 +MT 344 836 +MT 345 832 +MT 346 835 +MT 347 841 +MT 348 847 +MT 349 862 +MT 350 873 +MT 351 876 +MT 352 877 +MT 353 879 +MT 354 889 +MT 355 899 +MT 356 900 +MT 357 912 +MT 358 912 +MT 359 917 +MT 360 921 +MT 361 935 +MT 362 941 +MT 363 945 +MT 364 954 +MT 365 954 +MT 366 953 +MT 367 961 +MT 368 958 +MT 369 961 +MT 370 972 +MT 371 978 +MT 372 986 +MT 373 984 +MT 374 987 +MT 375 998 +MT 376 1002 +MT 377 1001 +MT 378 1006 +MT 379 1014 +MT 380 1008 +MT 381 1015 +MT 382 1021 +MT 383 1035 +MT 384 1040 +MT 385 1044 +MT 386 1056 +MT 387 1075 +MT 388 1091 +MT 389 1085 +MT 390 1084 +MT 391 1095 +MT 392 1101 +MT 393 1111 +MT 394 1123 +MT 395 1132 +MT 396 1149 +MT 397 1160 +MT 398 1168 +MT 399 1170 +MT 400 1181 +MT 401 1187 +MT 402 1184 +MT 403 1191 +MT 404 1200 +MT 405 1205 +MT 406 1196 +MT 407 1190 +MT 408 1192 +MT 409 1179 +MT 410 1168 +MT 411 1178 +MT 412 1180 +MT 413 1178 +MT 414 1185 +MT 415 1186 +MT 416 1184 +MT 417 1184 +MT 418 1182 +MT 419 1185 +MT 420 1182 +MT 421 1178 +MT 422 1178 +MT 423 1178 +MT 424 1171 +MT 425 1172 +MT 426 1173 +MT 427 1171 +MT 428 1167 +MT 429 1168 +MT 430 1171 +MT 431 1161 +MT 432 1177 +MT 433 1181 +MT 434 1180 +MT 435 1177 +MT 436 1188 +MT 437 1191 +MT 438 1176 +MT 439 1177 +MT 440 1164 +MT 441 1165 +MT 442 1170 +MT 443 1170 +MT 444 1169 +MT 445 1175 +MT 446 1178 +MT 447 1171 +MT 448 1167 +MT 449 1153 +MT 450 1149 +MT 451 1144 +MT 452 1138 +MT 453 1142 +MT 454 1130 +MT 455 1126 +MT 456 1115 +MT 457 1133 +MT 458 1145 +MT 459 1148 +MT 460 1147 +MT 461 1128 +MT 462 1140 +MT 463 1110 +MT 464 1111 +MT 465 1117 +MT 466 1117 +MT 467 1113 +MT 468 1120 +MT 469 1119 +MT 470 1107 +MT 471 1077 +MT 472 1078 +MT 473 1075 +MT 474 1080 +MT 475 1078 +MT 476 1076 +MT 477 1099 +MT 478 1100 +MT 479 1091 +MT 480 1097 +MT 481 1096 +MT 482 1104 +MT 483 1095 +MT 484 1089 +MT 485 1092 +MT 486 1090 +MT 487 1086 +MT 488 1066 +MT 489 1075 +MT 490 1093 +MT 491 1094 +MT 492 1091 +MT 493 1095 +MT 494 1095 +MT 495 1103 +MT 496 1144 +MT 497 1142 +MT 498 1145 +MT 499 1144 +MT 500 1083 +MT 501 1085 +MT 502 1091 +MT 503 1097 +MT 504 1106 +MT 505 1113 +MT 506 1118 +MT 507 1115 +MT 508 1103 +MT 509 1110 +MT 510 1117 +MT 511 1118 +MT 512 1120 +MT 513 1112 +MT 514 1108 +MT 515 1114 +MT 516 1115 +MT 517 1125 +MT 518 1120 +MT 519 1124 +MT 520 1127 +MT 521 1135 +MT 522 1130 +MT 523 1150 +MT 524 1152 +MT 525 1157 +MT 526 1142 +MT 527 1123 +MT 528 1139 +MT 529 1122 +MT 530 1126 +MT 531 1139 +MT 532 1128 +MT 533 1117 +MT 534 1121 +MT 535 1120 +MT 536 1121 +MT 537 1122 +MT 538 1115 +MT 539 1081 +MT 540 1077 +MT 541 1082 +MT 542 1078 +MT 543 1086 +MT 544 1085 +MT 545 1080 +MT 546 1065 +MT 547 1068 +MT 548 1076 +MT 549 1075 +MT 550 1079 +MT 551 1077 +MT 552 1093 +MT 553 1096 +MT 554 1093 +MT 555 1091 +MT 556 1093 +MT 557 1093 +MT 558 1094 +MT 559 1100 +MT 560 1099 +MT 561 1095 +MT 562 1064 +MT 563 1063 +MT 564 1064 +MT 565 1056 +MT 566 1055 +MT 567 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+MT 16514 926 +MT 16515 921 +MT 16516 910 +MT 16517 900 +MT 16518 883 +MT 16519 873 +MT 16520 877 +MT 16521 874 +MT 16522 864 +MT 16523 854 +MT 16524 844 +MT 16525 833 +MT 16526 826 +MT 16527 820 +MT 16528 813 +MT 16529 809 +MT 16530 798 +MT 16531 792 +MT 16532 786 +MT 16533 780 +MT 16534 771 +MT 16535 766 +MT 16536 763 +MT 16537 754 +MT 16538 753 +MT 16539 745 +MT 16540 726 +MT 16541 725 +MT 16542 720 +MT 16543 704 +MT 16544 695 +MT 16545 697 +MT 16546 692 +MT 16547 691 +MT 16548 690 +MT 16549 686 +MT 16550 678 +MT 16551 666 +MT 16552 651 +MT 16553 639 +MT 16554 629 +MT 16555 627 +MT 16556 622 +MT 16557 608 +MT 16558 598 +MT 16559 586 +MT 16560 581 +MT 16561 570 +MT 16562 558 +MT 16563 550 +MT 16564 542 +MT 16565 537 +MT 16566 525 +MT 16567 517 +MT 16568 508 +MT 16569 491 +MT 16570 487 +MT 16571 476 +MT 16572 465 +MT 16573 41 +MT 16574 32 +MT 16575 28 +MT 16576 24 +MT 16577 19 +MT 16578 7 diff --git a/workflows/mtdna_server_2.nf b/workflows/mtdna_server_2.nf index 9fb3bec..cb96eab 100644 --- a/workflows/mtdna_server_2.nf +++ b/workflows/mtdna_server_2.nf @@ -174,9 +174,14 @@ workflow MTDNA_SERVER_2 { INPUT_VALIDATION.out.excluded_ch ) + bams_filenames = bams_ch + .map (bam_file -> tuple(bam_file.name, file(bam_file))) + samples = INPUT_VALIDATION.out.mapping_ch .splitCsv(header: true, sep: '\t') - .map { row -> row.Sample } + .map { row -> tuple(row.Filename, row.Sample) } + .join(bams_filenames) + SAMPLE_REPORT( sample_report_file_ch, From 2d8ce941173c11ee6cd9c1abcbd41d7e5a7e3396 Mon Sep 17 00:00:00 2001 From: Lukas Forer Date: Sat, 2 Mar 2024 14:25:16 +0100 Subject: [PATCH 09/10] Fix issue with multiple contigs in coverage report --- modules/local/quality_control.nf | 4 +- modules/local/sample_report.nf | 10 ++++- reports/report.Rmd | 3 +- reports/sample.Rmd | 16 ++++++++ tests/mitocalling.nf.test.snap | 67 ++++++++++++++++++++++++++++++-- 5 files changed, 93 insertions(+), 7 deletions(-) diff --git a/modules/local/quality_control.nf b/modules/local/quality_control.nf index a084c5d..0010717 100644 --- a/modules/local/quality_control.nf +++ b/modules/local/quality_control.nf @@ -1,6 +1,6 @@ process QUALITY_CONTROL { - publishDir "${params.output_reports}", mode: "copy", pattern: '*.html' + publishDir "${params.output_reports}/multiqc", mode: "copy", pattern: '*.html' input: path excluded_samples @@ -10,6 +10,6 @@ process QUALITY_CONTROL { path "*.html" """ - multiqc . + multiqc . --filename index.html """ } \ No newline at end of file diff --git a/modules/local/sample_report.nf b/modules/local/sample_report.nf index b81e35e..e5d8f5d 100644 --- a/modules/local/sample_report.nf +++ b/modules/local/sample_report.nf @@ -27,10 +27,18 @@ process SAMPLE_REPORT { echo -e "Base Quality\t${params.baseQ}" >> params.txt echo -e "Map Quality\t${params.mapQ}" >> params.txt echo -e "Alignment Quality\t${params.alignQ}" >> params.txt + echo -e "Mutserv\t\${MUTSERV_VERSION}" >> params.txt + echo -e "Haplocheck\t\${HAPLOCHECK_VERSION}" >> params.txt + echo -e "Haplogrep\t\${HAPLOGREP_VERSION}" >> params.txt #TODO: split fwd and reverse? https://bioinformatics.stackexchange.com/questions/8649/how-can-i-calculate-coverage-at-single-bases-using-a-bam-file + + # Extract the value of Contig and store it in a variable + contig=`awk '\$2 == "Contig" {print \$3; exit}' "${statistics}"` + echo -e "Contig\tPosition\tCoverage" > coverage.tsv - samtools depth -a ${sample_bam_file} >> coverage.tsv + samtools index ${sample_bam_file} + samtools depth -a ${sample_bam_file} -r \$contig >> coverage.tsv Rscript -e "require('rmarkdown'); render('${report}', params = list( diff --git a/reports/report.Rmd b/reports/report.Rmd index 0164a8c..65c1ce4 100644 --- a/reports/report.Rmd +++ b/reports/report.Rmd @@ -4,7 +4,8 @@ output: flexdashboard::flex_dashboard: orientation: rows navbar: - - { title: "About", href: "https://mitoverse.i-med.ac.at/#!pages/contact", align: right } + - { title: "MultiQC", target: "_blank", href: "multiqc/index.html" } + - { title: "About", target: "_blank", href: "https://mitoverse.i-med.ac.at/#!pages/contact", align: right } params: haplogroups: ../tests/data/report/haplogroups.txt haplocheck: ../tests/data/report/haplocheck.txt diff --git a/reports/sample.Rmd b/reports/sample.Rmd index 482a734..2b1cfcf 100644 --- a/reports/sample.Rmd +++ b/reports/sample.Rmd @@ -191,6 +191,7 @@ variants %>% mutate( Variant_Label = link_to_variant(Pos, Ref, Variant) ) %>% + arrange(Pos, Ref, Variant) %>% select("Variant_Label", "VariantLevel", "Type_Label") %>% datatable( colnames=c("Variant", "Level", ""), @@ -206,3 +207,18 @@ variants %>% class = 'cell-border stripe' ) ``` + + +### Paramteres and Software Version + +```{r} +pipeline_params %>% + kable() +``` + + +--- + + +This report has been created with **[mtDNA-Server 2](https://mitoverse.i-med.ac.at)**, a nextflow pipeline developed by [Sebastian Schönherr](mailto:sebastian.schoenherr@i-med.ac.at), [Hansi Weissensteiner](mailto:hansi.weissensteiner@i-med.ac.at) and [Lukas Forer](mailto:lukas.forer@i-med.ac.at), Institute of Genetic Epidemiology, Medical University of Innsbruck. + \ No newline at end of file diff --git a/tests/mitocalling.nf.test.snap b/tests/mitocalling.nf.test.snap index 65bb23e..2b81aaa 100644 --- a/tests/mitocalling.nf.test.snap +++ b/tests/mitocalling.nf.test.snap @@ -3,18 +3,45 @@ "content": [ "variants.annotated.txt:md5,52dc3737a73bb80cf3a1c8946990c3e4" ], + "meta": { + "nf-test": "0.8.4", + "nextflow": "23.04.3" + }, "timestamp": "2024-03-02T13:11:34.901277407" }, + "Runs with 6 samples provided as testdata with mutect2": { + "content": [ + "variants.annotated.txt:md5,d5bcadb427d40ffc53a118bcd9bc5aae", + { + "tasksFailed": 0, + "tasksCount": 28, + "tasksSucceeded": 28 + } + ], + "meta": { + "nf-test": "0.8.4", + "nextflow": "23.04.3" + }, + "timestamp": "2024-03-02T14:11:30.627373" + }, "Runs with single BAM file and mutserve": { "content": [ "variants.annotated.txt:md5,ee00b39fc49e1f34e6affab00368181f" ], + "meta": { + "nf-test": "0.8.4", + "nextflow": "23.04.3" + }, "timestamp": "2024-03-02T13:09:45.954385792" }, "Runs with BAM file including different header contigs": { "content": [ "variants.annotated.txt:md5,a5345c7718757ded27246ae342b5f25b" ], + "meta": { + "nf-test": "0.8.4", + "nextflow": "23.04.3" + }, "timestamp": "2024-03-02T13:10:28.843431233" }, "Runs with two files and one goes through QC": { @@ -22,10 +49,44 @@ "variants.annotated.txt:md5,ef402762b03356a208583e434d96dfa3", { "tasksFailed": 0, - "tasksCount": 10, - "tasksSucceeded": 10 + "tasksCount": 12, + "tasksSucceeded": 12 + } + ], + "meta": { + "nf-test": "0.8.4", + "nextflow": "23.04.3" + }, + "timestamp": "2024-03-02T14:02:05.743122" + }, + "Runs with 6 samples provided as testdata with fusion": { + "content": [ + "variants.annotated.txt:md5,5043504edb7e0bf5492fafb63e64daa6", + { + "tasksFailed": 0, + "tasksCount": 40, + "tasksSucceeded": 40 + } + ], + "meta": { + "nf-test": "0.8.4", + "nextflow": "23.04.3" + }, + "timestamp": "2024-03-02T14:15:40.016064" + }, + "Runs with 6 samples provided as testdata with mutserve": { + "content": [ + "variants.annotated.txt:md5,d383a9cb2a1f46c96770390c61002bfc", + { + "tasksFailed": 0, + "tasksCount": 30, + "tasksSucceeded": 30 } ], - "timestamp": "2024-03-02T13:12:29.64633988" + "meta": { + "nf-test": "0.8.4", + "nextflow": "23.04.3" + }, + "timestamp": "2024-03-02T14:08:07.908258" } } \ No newline at end of file From 2c7671c2210200584cb959df2ac10e482dbfd683 Mon Sep 17 00:00:00 2001 From: Lukas Forer Date: Sat, 2 Mar 2024 15:05:43 +0100 Subject: [PATCH 10/10] Sort duplicate variants by ref and alt --- modules/local/merging_variants.nf | 2 +- tests/mitocalling.nf.test.snap | 24 ++++++++++++------------ 2 files changed, 13 insertions(+), 13 deletions(-) diff --git a/modules/local/merging_variants.nf b/modules/local/merging_variants.nf index f58a2f4..66bcb9f 100644 --- a/modules/local/merging_variants.nf +++ b/modules/local/merging_variants.nf @@ -14,7 +14,7 @@ process MERGING_VARIANTS { csvtk sort \ -t variants.concat.txt \ - -k ID:N -k Pos:n -k Type:nr \ + -k ID:N -k Pos:n -k Ref:N -k Variant:N -k Type:nr \ -T -o variants.sorted.txt if [[ ${mode} == "fusion" ]] diff --git a/tests/mitocalling.nf.test.snap b/tests/mitocalling.nf.test.snap index 2b81aaa..ba56425 100644 --- a/tests/mitocalling.nf.test.snap +++ b/tests/mitocalling.nf.test.snap @@ -1,17 +1,17 @@ { "Runs with single BAM file and mutect2": { "content": [ - "variants.annotated.txt:md5,52dc3737a73bb80cf3a1c8946990c3e4" + "variants.annotated.txt:md5,7dc0d48ba82f76bca62ed2dddbf7d7bb" ], "meta": { "nf-test": "0.8.4", "nextflow": "23.04.3" }, - "timestamp": "2024-03-02T13:11:34.901277407" + "timestamp": "2024-03-02T14:55:31.73871" }, "Runs with 6 samples provided as testdata with mutect2": { "content": [ - "variants.annotated.txt:md5,d5bcadb427d40ffc53a118bcd9bc5aae", + "variants.annotated.txt:md5,1e00242adc88c7cb44adea0dd77c325e", { "tasksFailed": 0, "tasksCount": 28, @@ -22,17 +22,17 @@ "nf-test": "0.8.4", "nextflow": "23.04.3" }, - "timestamp": "2024-03-02T14:11:30.627373" + "timestamp": "2024-03-02T15:00:48.608014" }, "Runs with single BAM file and mutserve": { "content": [ - "variants.annotated.txt:md5,ee00b39fc49e1f34e6affab00368181f" + "variants.annotated.txt:md5,77be10aad2e2ba4fc50296b693e1d8e0" ], "meta": { "nf-test": "0.8.4", "nextflow": "23.04.3" }, - "timestamp": "2024-03-02T13:09:45.954385792" + "timestamp": "2024-03-02T14:53:52.048799" }, "Runs with BAM file including different header contigs": { "content": [ @@ -46,7 +46,7 @@ }, "Runs with two files and one goes through QC": { "content": [ - "variants.annotated.txt:md5,ef402762b03356a208583e434d96dfa3", + "variants.annotated.txt:md5,fb9b8ca4410e67f47bb2f0e4c348f908", { "tasksFailed": 0, "tasksCount": 12, @@ -57,11 +57,11 @@ "nf-test": "0.8.4", "nextflow": "23.04.3" }, - "timestamp": "2024-03-02T14:02:05.743122" + "timestamp": "2024-03-02T14:56:19.810049" }, "Runs with 6 samples provided as testdata with fusion": { "content": [ - "variants.annotated.txt:md5,5043504edb7e0bf5492fafb63e64daa6", + "variants.annotated.txt:md5,3a0f3b225f5618d8a3c0d7013911f876", { "tasksFailed": 0, "tasksCount": 40, @@ -72,11 +72,11 @@ "nf-test": "0.8.4", "nextflow": "23.04.3" }, - "timestamp": "2024-03-02T14:15:40.016064" + "timestamp": "2024-03-02T15:04:51.140209" }, "Runs with 6 samples provided as testdata with mutserve": { "content": [ - "variants.annotated.txt:md5,d383a9cb2a1f46c96770390c61002bfc", + "variants.annotated.txt:md5,c2f723faf745820f9767548dc52ee39a", { "tasksFailed": 0, "tasksCount": 30, @@ -87,6 +87,6 @@ "nf-test": "0.8.4", "nextflow": "23.04.3" }, - "timestamp": "2024-03-02T14:08:07.908258" + "timestamp": "2024-03-02T14:57:46.95114" } } \ No newline at end of file