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Pathogen infection is still a huge problem threatening human health all over the world, and it has brought a heavy burden to clinical diagnosis and treatment
.
The traditional clinical pathogen identification is mainly based on culture and biochemical detection, but with the continuous advancement of next-generation sequencing technology, metagenomic sequencing ( mNGS ) greatly improves the efficiency of pathogen detection and helps to identify difficult-to-
At present, mNGS mainly targets the DNA of pathogenic microorganisms and uses second-generation sequencing technology to generate shorter sequences.
Therefore, some pathogenic microorganisms whose main genetic material is RNA are ignored .
Even though RNA- based analysis or metatranscriptome research is relatively rare, they all show that targeted RNA detection can effectively reveal functionally active members / genes; in addition, although the bacterial genome has only a few rDNA copies, the content of rRNA molecules is active However, it is extremely high in cells.
In theory, targeted RNA can avoid the predominant but dormant microorganisms accounting for the proportion of sequencing reads .
The third-generation sequencing represented by Oxford Nanopore Technology's sequencing technology has the advantages of rapid library preparation and ultra-long read length, which can effectively improve the detection of pathogens in clinical samples
.
At the same time, ONT direct RNA sequencing technology also provides the possibility for unbiased detection of RNA pathogens in
Studies have shown that compared with mNGS , mtNGS increases the ratio of microbial readings, facilitates the identification of bacterial pathogens, and can detect fungi, viruses and antibiotic resistance genes, which are basically consistent with traditional clinical test results
.
In addition, the ONT- based mtTGS further improves the identification of pathogenic microorganisms due to its longer read length, and also speeds up the diagnosis time
The above research results were published in the journal Advanced Science in October 2021.
Assistant researcher Zhao Na, Ph.
D student Cao Jiabao, Ph.
Full text link: https://onlinelibrary.
wiley.
com/doi/10.
