Pathogen Real-Time Identification by Sequencing
Pathogen Identification Directly from Whole Blood within 24 Hours !
Pathogen Real-Time Identification by Sequencing , is a molecular diagnostic technique that combines real-time PCR (polymerase chain reaction) with next-generation sequencing (NGS) to rapidly identify and characterize pathogens present in a given sample.
mNGS in clinical microbiology laboratories :
Metagenomic next-generation sequencing (mNGS) is increasingly being applied in clinical laboratories for unbiased culture-independent diagnosis. Whether it can be a next routine pathogen identification tool has become a topic of concern...
The steps of this method :
1. Sample collection : A sample, such as a clinical specimen or an environmental sample, is collected from the suspected source of infection or contamination.
2. DNA or RNA extraction: The genetic material (DNA or RNA) is extracted from the sample using standard laboratory protocols. This step isolates the pathogen's genetic material for subsequent analysis.
3. Real-time PCR: Real-time PCR is performed to amplify specific regions of the pathogen's genetic material. This technique uses primers and probes that are designed to target conserved regions within the pathogen's genome. Real-time PCR allows for the amplification and quantification of the pathogen's DNA or RNA in the sample.
4. Library preparation: The amplified DNA or RNA from the real-time PCR step is then used to generate a sequencing library. This involves preparing the DNA or RNA fragments for sequencing by adding adapter sequences and incorporating indexes or barcodes to enable sample multiplexing.
5. Next-generation sequencing: The prepared library is subjected to NGS using high-throughput sequencing platforms. The sequencing generates millions of short DNA or RNA sequences, which represent the genetic material of the pathogen and other organisms present in the sample.
6. Data analysis: The sequence data obtained from NGS is analyzed using bioinformatics tools and algorithms. This involves comparing the obtained sequences to known pathogen databases or reference genomes to identify the specific pathogens present in the sample. The analysis can also provide information about the genetic variation, virulence factors, and drug resistance profiles of the identified pathogens.
Advantages of PaRTI-Seq™ :
It offers several advantages in the field of pathogen detection and identification. Here are some key advantages of PaRTI-Seq™ :
- Rapid and comprehensive pathogen identification.
- High sensitivity and specificity.
- Simultaneous detection of multiple pathogens.
- Potential for drug resistance profiling.
- Unbiased detection and discovery.
- Potential for epidemiological investigations.