To perform an E-BURST analysis of three sequence types (STs) of Klebsiella aerogenes, follow these steps: ### Step 1: Obtain MLST Data 1. **Collect MLST Data**: Ensure you have the MLST profiles for the three STs of Klebsiella aerogenes. Each ST is defined by the alleles at seven housekeeping gene loci. 2. **Database Access**: You can obtain MLST data from public databases such as the PubMLST database or by sequencing the housekeeping genes of your isolates. ### Step 2: Prepare Data for E-BURST 1. **Format Data**: Convert the MLST data into a format suitable for E-BURST analysis. Typically, this involves creating a text file with the allele profiles for each ST. 2. **Example Format**: ``` ST gene1 gene2 gene3 gene4 gene5 gene6 gene7 1 3 5 7 9 11 13 15 2 3 5 7 9 12 13 15 3 4 5 7 9 11 13 15 ``` ### Step 3: Perform E-BURST Analysis 1. **Access E-BURST Tool**: Use the E-BURST tool available online or download the software from the E-BURST website. 2. **Input Data**: Upload your formatted MLST data into the E-BURST tool. 3. **Run Analysis**: Execute the E-BURST analysis to identify clonal complexes and patterns of evolutionary descent among the STs. ### Step 4: Interpret Results 1. **Clonal Complexes**: E-BURST will group STs into clonal complexes based on their allelic profiles. STs that differ at only one of the seven loci are considered single-locus variants (SLVs) and are grouped together. 2. **Founders**: Identify the founder of each clonal complex, which is the ST with the largest number of single-locus variants. 3. **Patterns**: Analyze the patterns of descent and relatedness among the STs to understand the evolutionary relationships and potential transmission dynamics. ### Example Interpretation - **ST1 and ST2**: If ST1 and ST2 are SLVs, they will be grouped together in the same clonal complex, indicating a close evolutionary relationship. - **ST3**: If ST3 is a double-locus variant (DLV) of ST1, it will be linked to ST1 in the E-BURST diagram, showing a more distant relationship. ### Additional Resources - **Kleborate Tool**: For a more comprehensive analysis, you can use the Kleborate tool, which is designed for analyzing genomes of Klebsiella species, including Klebsiella aerogenes. - **MLST Databases**: Access MLST databases to obtain and verify the allele profiles of your STs. By following these steps, you can effectively perform an E-BURST analysis to understand the evolutionary relationships and clonal structure of the three STs of Klebsiella aerogenes.