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Multi-Locus Sequence Typing

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Overview

Multi-locus sequence typing (MLST) is a molecular biology method with high resolution based on the analysis of internal fragments of multiple housekeeping genes by PCR and sequencing. It can be used to identify and classify unidentified bacterial and fungal organisms at the species and strain levels. It has been applied in both molecular epidemiological studies and molecular evolution analysis. We provide MLST service to help you characterize isolates of microbial species and to accelerate studies of SNPs, resistance genes, and virulence factors of strains.

Our Advantages:
  • Provide comprehensive services for microbial identification.
  • Provide exquisite gel electropherogram.
  • High-resolution and high-reproducibility data.
  • Extensive experience in handling various samples.
Tell Us About Your Project

We are dedicated to providing outstanding customer service and being reachable at all times.

Request a Quote

Introduction to our multi-locus sequence typing platform

Our MLST platform allows the characterization of microbial species/strains by analyzing the internal fragments of 6-8 housekeeping genes (about 450 bp) that can provide acceptable discriminatory power. The different sequences of these housekeeping genes are assigned as alleles. Alleles are initially amplified by PCR and analyzed by Sanger sequencing or next-generation sequencing (NGS). The reads are then aligned before compared by phylogenetic approaches to distinguish the difference among individuals and clades. This technology does not require live microbes or high-quality genomic DNA, so the results are not easily affected by sample transport and manual manipulation. This analysis can directly analyze microbes that cannot be isolated and grown in pure culture.

MLST has several advantages over other typing approaches. First, compared to phenotypic approaches, it can detect changes at the DNA level. Second, standardized sequencing methods yield accurate and highly sensitive sequence results compared to fragment-based methods. Our MLST service can be used to not only compare the diversity of the alleles of the strains, but also study the resistant mechanism of drug-resistant strains, specific genotypes related to virulence or antigen, disease prevalence caused by new variants, biological evolution, and population structure. Our MLST platform can support multiple fields, including scientific research, agriculture, pharmaceutical industry, etc.

Multi-locus sequence typing service workflow

Bioinformatics Analysis

  1. Align sequences
  2. Identify polymorphic sites
  3. Characterize isolates of bacteria, fungi, and mycoplasma, etc.
  4. Construct phylogenetic trees
  5. Determine the rate of substitution at synonymous and non-synonymous sites (dN and dS, respectively).

Sample Requirement

    1. Microbial genomic DNA: total amount > 500 ng, concentration > 10 ng/ul.
    2. 1.8 < OD260/280 < 2.0, no degradation or contamination.

Sampling kits: We provide a range of microbial sampling kits for clients, including MicroCollect™ oral sample microbial collection products and MicroCollect™ stool sample collection products.

Deliverables: The experimental report includes information on experimental methods, primers, PCR conditions, gel electropherogram, MLST allele and sequence type (ST), phylogenetic tree, etc.

References

  1. Maiden M C J, et al. Multilocus sequence typing: a portable approach to the identification of clones within populations of pathogenic microorganisms. Proceedings of the National Academy of Sciences, 1998, 95(6): 3140-3145.
  2. Boonsilp S, et al. A single multilocus sequence typing (MLST) scheme for seven pathogenic Leptospira species. PLoS neglected tropical diseases, 2013, 7(1): e1954.
* For Research Use Only. Not for use in diagnostic procedures or other clinical purposes.



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