Making a Long Story Short: Quick Bacterial Diagnostics with High-Molecular Weight DNA
In this webinar, you will learn about:
- The importance of HMW DNA in pathogen genomics
- Application of long-read sequencing in bacterial diagnostics
- Implementation of genomics in innovative all-in-one bacterial diagnostics
Bacterial diagnostics in human and veterinary medicine relies majorly on bacterial cultures or targeted short amplicon PCR-based tests. While culture-based assays are time-consuming or restricted for various important species (up to 2 weeks for Mycoplasma spp.), molecular assays provide a quick output. Hence, combined approaches are required to deliver answers on species identification, typing & antimicrobial susceptibility testing (AST). Making these long diagnostic stories short has been facilitated by the release of new affordable and real-time sequencing approaches (e.g. Oxford Nanopore Technologies). Exploiting long-read sequencing to the most, requires the generation of High-Molecular Weight (HMW) nucleic acids.
Rapid MinION sequencing could be performed on various bacterial species using HMW DNA extracts from the Wizard® Genomic DNA purification kit (Promega). This allowed us to collect more comprehensive data in real-time for faster species identification, typing & verification of the presence of virulence factor genes and antimicrobial resistance mediators. In addition, implementation of longer bacterial reads (N50 = ±20kbp) required less sequencing time to obtain 100x covered complete & high-quality bacterial genomes from datasets of Gram-positive, Gram-negative & Mycoplasma spp. bacteria.
In conclusion, longer DNA molecules allow to shorten a long diagnostics workflow by quickly generating complete & high-quality bacterial genomes in an all-in-one sequencing-based bacterial diagnostic tests in human and veterinary medicine.
Research & Development Scientist
Ruchika Sharma, PhD
Product Specialist - Genomic Solutions
Ruchika Sharma is a Product Specialist in the Genomic Solutions Group at Promega Corporation. She manages a portfolio of Promega’s sample preparation products for in-vitro diagnostics and manual purification and works with customers in different market segments globally to understand and address their unmet sample preparation needs. She also works with Promega’s cross-functional teams to provide solutions tailored to customer needs for any specific applications of Promega’s products, including the Wizard® HMW DNA Extraction Kit for isolation of high-molecular weight DNA for long read sequencing. Ruchika holds a Ph.D. in Biochemistry and Molecular Biology from the Indian Institute of Science and did her postdoctoral training at the National Institutes of Health and the University of Wisconsin-Madison, focusing on the role of molecular chaperones in protein folding and human diseases.