#include #include #include #include #include #include #include #include #include "nnls.h" #include "kmer_utils.h" #include "quikr_functions.h" #include "quikr.h" #define sensing_matrix(i,j) (sensing_matrix[width*i + j]) #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-f, --sensing-fasta\n\tlocation of the fasta file database used to create the sensing matrix (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 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-fasta", required_argument, 0, 'f'}, {"sensing-matrix", required_argument, 0, 's'}, {"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:f:s:i:o:hdvV", long_options, &option_index); if (c == -1) break; switch (c) { case 'k': kmer = atoi(optarg); break; case 'l': lambda = atoi(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); break; case 'h': printf("%s\n", USAGE); exit(EXIT_SUCCESS); break; 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(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(4, kmer); width = width + 1; // load counts matrix unsigned long long *integer_counts = get_kmer_counts_from_file(input_fasta_filename, kmer); double *count_matrix = malloc(sizeof(double) * width); if(count_matrix == NULL) { fprintf(stderr, "Could not allocate memory:\n"); exit(EXIT_FAILURE); } count_matrix[0] = 0; for(x = 1; x < width; x++) count_matrix[x] = (double)integer_counts[x-1]; free(integer_counts); // normalize our count_matrix normalize_matrix(count_matrix, 1, width); for(x = 0; x < width; x++) count_matrix[x] = count_matrix[x] * lambda; // load sensing matrix struct matrix *sensing_matrix = load_sensing_matrix(sensing_matrix_filename); if(sensing_matrix->kmer != kmer) { fprintf(stderr, "The sensing_matrix was trained with a different kmer than your requested kmer\n"); exit(EXIT_FAILURE); } // multiply our sensing matrix by lambda for(x = 1; x < sensing_matrix->sequences; x++) { for(y = 0; y < width - 1; y++) { sensing_matrix->matrix[width*x + y] = sensing_matrix->matrix[width*x + y] * lambda; } } // set the first column of our sensing matrix to 0 for(x = 0; x < sensing_matrix->sequences; x++) { sensing_matrix->matrix[width * x] = 1.0; } // run NNLS double *solution = nnls(sensing_matrix->matrix, count_matrix, sensing_matrix->sequences, 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); return EXIT_SUCCESS; }