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AIM:To identify and assess the novel makers for detection of Shiga toxin producing Escherichia coli (STEC) O157:H7 with an integrated computational and experimental approach. METHODS:High-throughput NCBI blast (E-value cutoff e-5) was used to search homologous genes among all sequenced prokaryotic genomes of each gene encoded in each of the three strains of STEC O157:H7 with complete genomes,aiming to find unique genes in O157:H7 as its potential markers. To ensure that the identified markers from the three strains of STEC O157:H7 can serve as general markers for all the STEC O157:H7 strains,a genomic barcode approach was used to select the markers to minimize the possibility of choosing a marker gene as part of a transposable element. Effectiveness of the markers predicted was then validated by running polymerase chain reaction (PCR) on 18 strains of O157:H7 with 5 additional genomes used as negative controls. RESULTS:The blast search identified 20,16 and 20 genes,respectively,in the three sequenced strains of STEC O157:H7,which had no homologs in any of the other prokaryotic genomes. Three genes,wzy,Z0372 and Z0344,common to the three gene lists,were selected based on the genomic barcode approach. PCR showed an identification accuracy of 100% on the 18 tested strains and the 5 controls. CONCLUSION:The three identified novel markers,wzy,Z0372 and Z0344,are highly promising for the detection of STEC O157:H7,in complementary to the known markers.
AIM: To identify and evaluate the novel makers for detection of Shiga toxin producing Escherichia coli (STEC) O157: H7 with an integrated computational and experimental approach. METHODS: High-throughput NCBI blast (E-value cutoff e-5) was used to search homologous genes among all sequenced prokaryotic genomes of each of the three genes encoded in each of the three strains of STEC O157: H7 with complete genomes, aiming to find unique genes in O157: H7 as its potential markers. The STEC O157: H7 can serves as general markers for all the STEC O157: H7 strains, a genomic barcode approach was used to select the markers to minimize the possibility of choosing a marker gene as part of a transposable element. Effectiveness of the markers predicted was then validated by running polymerase chain reaction (PCR) on chromosome 18 of O157: H7 with 5 additional genomes used as negative controls. RESULTS: The blast search identified 20, 16 and 20 genes, respectively, in the Three sequenced strains of STEC O157: H7, which had no homologs in any of the other prokaryotic genomes. Three genes, wzy, Z0372 and Z0344, common to the three gene lists, were selected based on the genomic barcode approach. PCR showed an identification Accuracy of 100% on the 18 tested strains and the 5 controls. CONCLUSION: The three identified novel markers, wzy, Z0372 and Z0344 are are promising for the detection of STEC O157: H7, in complementary to the known markers.