Development of diagnostic method for comprehensive viral detection using not so random primer sets

2014 Fiscal Year Final Research Report

• Project/Area Number: 25670271
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Laboratory medicine
• Research Institution: Osaka University
• Principal Investigator: NAKAMURA SHOTA 大阪大学, 微生物病研究所, 助教 (90432434)
• Project Period (FY): 2013-04-01 – 2015-03-31
• Keywords: 感染症 / メタゲノム

Outline of Final Research Achievements

Unbiased next-generation sequencing approaches enable a diagnostic method for comprehensive pathogen detection. This method employs a single common protocol without specific treatment for each pathogen, which consists of the amplification of DNA/RNA in clinical specimens by a random primer set, metagenomic shotgun sequencing, and identification of pathogenic genes. Our previous studies have revealed that a large amount of human genomic DNA in clinical specimens decreases the sensitivity for pathogen detection. Therefore, we tried to develop the subtraction method for human-derived sequences using not so random primer sets. Using the newly designed primer sets, we were able to subtract the human sequences and amplify the target viral sequences from the specimen of influenza A infection. However, undesirable environmental bacterial sequences were obtained coincidentally. Further study on improving the sensitivity by redesign of the primer sets and protocols needs to be considered.

Free Research Field

病態検査学