ChapterDx MLSTnext Hepatitis A Genotyping NGS Assay
ChapterDx MLSTnext Hepatitis A genotyping NGS assay analyzes 10 highly polymorphic loci spanning Hepatitis A virus genome. The combination of the highly polymorphic information at these loci provides high-resolution genotype information, which also enables tracking-and-tracing the source of infection.
Description
ChapterDx MLSTnext Hepatitis A genotyping NGS assay is a unique and highly multiplex PCR assay that amplifies and analyzes 10 highly polymorphic loci spanning Hepatitis A virus genome. The assay is a single-tube and single-step PCR reaction, where amplification and indexing take places simultaneously in the same reaction. This assay provides high-resolution genotype information for Hepatitis A virus and can detect co-presence of different genotypes within same sample. The high-resolution results enables identification of source of infection during Hepatitis A outbreaks. ChapterDx Hepatitis A data analysis software is included.
Technology
ChapterDx Hepatitis A Genotyping NGS assay consists of the following steps:
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RNA extraction (not provided)
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Multiplex RT-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 RT-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 less than half hour to complete data analysis.
Features
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Sequence analysis of 10 highly polymorphic loci across Hepatitis A virus genome
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Provides high-resolution Hepatitis A genotyping
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Detection of multiple genotypes/subtypes co-exisitng in the same sample
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Excellent tool for tracking-and-tracing the source of infection during outbreaks
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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 to high-throughput scales
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Data analysis software included
Location of Amplicons in Hepatitis A Genome

ChapterDx HAV Genotyping NGS Assay Workflow
