#include #include #include #include #include #include #include #include "nnls.h" #include "kmer_utils.h" #include "quikr_functions.h" #include "quikr.h" #define USAGE "Usage:\n\tquikr [OPTION...] - Calculate estimated frequencies of bacteria in a sample.\n\nOptions:\n\n-i, --input\n\tthe sample's fasta file of NGS READS (fasta format)\n\n-s, --sensing-matrix\n\t location of the sensing matrix. (trained from quikr_train)\n\n-k, --kmer\n\tspecify what size of kmer to use. (default value is 6)\n\n-l, --lambda\n\tlambda value to use. (default value is 10000)\n\n-o, --output\n\tOTU_FRACTION_PRESENT a vector representing the percentage of database sequence's presence in sample. (csv output)\n\n-v, --verbose\n\tverbose mode.\n\n-V, --version\n\tprint version." int main(int argc, char **argv) { int c; char *input_fasta_filename = NULL; char *sensing_matrix_filename = NULL; char *output_filename = NULL; unsigned long long x = 0; unsigned long long y = 0; unsigned long long z = 0; unsigned long long rare_value = 0; unsigned long long rare_width = 0; double rare_percent = 1.0; unsigned long long width = 0; unsigned int kmer = 6; unsigned long long lambda = 10000; int verbose = 0; while (1) { static struct option long_options[] = { {"input", required_argument, 0, 'i'}, {"kmer", required_argument, 0, 'k'}, {"lambda", required_argument, 0, 'l'}, {"output", required_argument, 0, 'o'}, {"sensing-matrix", required_argument, 0, 's'}, {"rare-percent", required_argument, 0, 'r'}, {"verbose", no_argument, 0, 'v'}, {"version", no_argument, 0, 'V'}, {"help", no_argument, 0, 'h'}, {"debug", no_argument, 0, 'd'}, {0, 0, 0, 0} }; int option_index = 0; c = getopt_long (argc, argv, "k:l:s:r:i:o:r:hdvV", long_options, &option_index); if (c == -1) break; switch (c) { case 'k': kmer = atoi(optarg); break; case 'l': lambda = atoi(optarg); break; case 'r': rare_percent = atof(optarg); break; case 's': sensing_matrix_filename = optarg; break; case 'i': input_fasta_filename = optarg; break; case 'o': output_filename = optarg; break; case 'v': verbose = 1; break; case 'V': printf("%s\n", VERSION); exit(EXIT_SUCCESS); case 'h': printf("%s\n", USAGE); exit(EXIT_SUCCESS); default: break; } } if(sensing_matrix_filename == NULL) { fprintf(stderr, "Error: sensing matrix filename (-s) must be specified\n\n"); fprintf(stderr, "%s\n", USAGE); exit(EXIT_FAILURE); } if(output_filename == NULL) { fprintf(stderr, "Error: output filename (-o) must be specified\n\n"); fprintf(stderr, "%s\n", USAGE); exit(EXIT_FAILURE); } if(input_fasta_filename == NULL) { fprintf(stderr, "Error: input fasta file (-i) must be specified\n\n"); fprintf(stderr, "%s\n", USAGE); exit(EXIT_FAILURE); } if(rare_percent <= 0 || rare_percent > 1.0) { fprintf(stderr, "Error: rare percent must be between 0 and 1\n"); exit(EXIT_FAILURE); } if(verbose) { printf("kmer: %u\n", kmer); printf("lambda: %llu\n", lambda); printf("fasta: %s\n", input_fasta_filename); printf("sensing matrix: %s\n", sensing_matrix_filename); printf("output: %s\n", output_filename); } if(access (sensing_matrix_filename, F_OK) == -1) { fprintf(stderr, "Error: could not find %s\n", sensing_matrix_filename); exit(EXIT_FAILURE); } if(access (input_fasta_filename, F_OK) == -1) { fprintf(stderr, "Error: could not find %s\n", input_fasta_filename); exit(EXIT_FAILURE); } if(kmer == 0) { fprintf(stderr, "Error: zero is not a valid kmer\n"); exit(EXIT_FAILURE); } // 4 "ACGT" ^ Kmer gives us the size of output rows width = pow_four(kmer); // load sensing matrix struct matrix *sensing_matrix = load_sensing_matrix(sensing_matrix_filename, kmer); if(verbose) { printf("width: %llu\n", width); printf("sequences: %llu\n", sensing_matrix->sequences); } // load counts matrix double *count_matrix = malloc(width * sizeof(double)); check_malloc(count_matrix, NULL); // convert our matrix into doubles { unsigned long long *integer_counts = get_kmer_counts_from_file(input_fasta_filename, kmer); for(x = 0; x < width; x++) { count_matrix[x] = (double)integer_counts[x]; } free(integer_counts); } // get_rare_value get_rare_value(count_matrix, width, rare_percent, &rare_value, &rare_width); if(verbose) printf("there are %llu values less than %llu\n", rare_width, rare_value); // add a extra space for our zero's array, so we can set the first column to 1's rare_width++; // store our count matrix double *count_matrix_rare = calloc(rare_width, sizeof(double)); check_malloc(count_matrix_rare, NULL); double *sensing_matrix_rare = calloc(rare_width * sensing_matrix->sequences, sizeof(double)); check_malloc(sensing_matrix_rare, NULL); // copy only kmers from our original counts that match our rareness percentage // in both our count matrix and our sensing matrix // // y = 1 because we are offsetting the arrah by 1, so we can set the first row to all 1's for(x = 0, y = 1; x < width; x++) { if(count_matrix[x] <= rare_value) { count_matrix_rare[y] = count_matrix[x]; for(z = 0; z < sensing_matrix->sequences; z++) sensing_matrix_rare[z*rare_width + y] = sensing_matrix->matrix[z*width + x]; y++; } } // normalize our kmer counts and our sensing_matrix normalize_matrix(sensing_matrix_rare, sensing_matrix->sequences, rare_width); normalize_matrix(count_matrix_rare, 1, rare_width); // multiply our kmer counts and sensing matrix by lambda for(x = 0; x < sensing_matrix->sequences; x++) { for(y = 1; y < rare_width; y++) { sensing_matrix_rare[rare_width*x + y] *= lambda; } } for(x = 0; x < sensing_matrix->sequences; x++) { sensing_matrix_rare[x*rare_width] = 1.0; } // count_matrix's first element should be zero count_matrix_rare[0] = 0; for(x = 1; x < rare_width; x++) count_matrix_rare[x] *= lambda; double *solution = nnls(sensing_matrix_rare, count_matrix_rare, sensing_matrix->sequences, rare_width); // normalize our solution vector normalize_matrix(solution, 1, sensing_matrix->sequences); // output our matrix FILE *output_fh = fopen(output_filename, "w"); if(output_fh == NULL) { fprintf(stderr, "Could not open %s for writing\n", output_filename); exit(EXIT_FAILURE); } for(x = 0; x < sensing_matrix->sequences; x++) fprintf(output_fh, "%.10lf\n", solution[x]); fclose(output_fh); free(solution); free(count_matrix); free(sensing_matrix); free(count_matrix_rare); free(sensing_matrix_rare); return EXIT_SUCCESS; }