One-step multiplex reverse-transcriptase PCR for detecting pandemic (H1N1) 2009 influenza virus

Area of Science:

• Veterinary Virology
• Molecular Diagnostics
• Public Health

Background:

• The 2009 pandemic H1N1 influenza virus emerged in swine and spread globally, causing significant human mortality.
• Early identification and monitoring of the virus in animal populations, particularly swine, are crucial for pandemic control.

Purpose of the Study:

• To develop and evaluate a sensitive diagnostic assay for distinguishing pandemic (H1N1) 2009 influenza virus from endemic swine influenza viruses.
• To provide a reliable tool for routine monitoring of pandemic (H1N1) 2009 influenza virus in pig herds.

Main Methods:

• Development of a one-step multiplex reverse transcriptase polymerase chain reaction (RT-PCR) assay targeting the matrix gene.
• Validation of the assay's sensitivity using in vitro-transcribed target RNA and virus cultures.
• Comparison of the assay's performance with existing influenza detection methods.

Main Results:

• The developed RT-PCR assay demonstrated high sensitivity, detecting as little as 100 copies of target RNA or 0.01 TCID(50)/ml of virus.
• The assay successfully discriminated pandemic (H1N1) 2009 influenza virus from endemic swine influenza strains.
• Performance was comparable to conventional and real-time RT-PCR assays for pandemic (H1N1) 2009 influenza detection.

Conclusions:

• The one-step multiplex RT-PCR assay is a sensitive and specific tool for identifying pandemic (H1N1) 2009 influenza virus in swine.
• This assay can be effectively utilized for regular surveillance of pandemic (H1N1) 2009 influenza virus in pig populations.
• Regular monitoring of swine herds is essential for controlling the spread of pandemic influenza.