ChapterDx MLSTnext Campylobacter Serotyping NGS Assay
ChapterDx MLSTnext Campylobacter serotyping NGS assay analyzes 80 most polymorphic loci spanning C. jejuni, C. Lari, C. coli, and C. fetus genomes. The combination of sequences at those loci enables high-resolution serotyping. In addition, this assay is able to detect more than one Campylobacter species from the same sample.
Description
ChapterDx MLSTnext Campylobacter serotyping NGS assay is a highly multiplex PCR assay that amplifies and analyzes 80 most polymorphic loci spanning the entire C. jejuni, C. Lari, C. coli and C. fetus genomes. The assay is a single-tube and single-step PCR reaction, where amplification and indexing take places simultaneously in the same reaction. The assay can be used for detection, serotyping, subtyping, identification of multiple serotypes in the same sample, and surveillance. The assay comes with data analysis software.
Technology
ChapterDx MLSTnext Campylobacter Serotyping NGS assay consists of the following steps:
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DNA extraction (not provided)
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Multiplex PCR
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Pooling and purification
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Next-generation sequencing
In the PCR reaction, both amplification and indexing/barcoding of each specimen occurs simultaneously. After PCR reaction, amplicons for different samples are pooled in one tube. The following procedures deal with only one tube regardless of sample numbers. After the cleanup step, the library is sequenced on an Illumina sequencing platform. The entire workflow takes about 24 hours, of which about 3 hours is benchwork/incubation time and 17 hours of sequencing. It takes about half hour to complete data analysis.
Features
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Analyze sequences for 80 polymorphic loci across Campylobacter species genomes
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Detect more than one Campylobacter species from the same sample
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Covers C. jejuni. C. lari, C. coli, and C. fetus genomes (including C. avium, C. gracilis and C. ureolyticus)
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Surveillance and tracking-and-tracing the source of contamination
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Single-tube and single-step PCR
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User-friendly workflow and easy to automate
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Cost-efficient, and suitable for low and high-throughput sample scales
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Data analysis software included
ChapterDx MLSTnext Campylobacter Serotyping NGS Assay Workflow
