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United States Department of Agriculture

Agricultural Research Service

Research Project: SWINE VIRAL DISEASES PATHOGENESIS AND IMMUNOLOGY Title: One Step Real-Time RT-PCR for 2009 Pandemic H1N1 Matrix Gene Detection and Quantitation in Clinical Samples

Authors
item Lorusso, Alessio
item Faaberg, Kay
item Killian, Mary Lea -
item Koster, Leo -
item Vincent, Amy

Submitted to: American Association of Swine Veterinarians Annual Meeting
Publication Type: Abstract Only
Publication Acceptance Date: October 1, 2009
Publication Date: March 6, 2010
Citation: Lorusso, A., Faaberg, K.S., Killian, M., Koster, L., Vincent, A.L. 2010. One Step Real-Time RT-PCR for 2009 Pandemic H1N1 Matrix Gene Detection and Quantitation in Clinical Samples [abstract]. American Association of Swine Veterinarians. p. 369.

Technical Abstract: In the spring of 2009, a novel H1N1 influenza A virus began to spread among humans worldwide. The genomic features of the new pandemic H1N1 were immediately identified: it contained gene segments with ancestors in North American and Eurasian swine influenza virus (SIV) lineages providing the virus a unique genetic constellation as well as genomic markers. Worldwide, the new H1N1 has not been demonstrated to be circulating among pigs and human infection was not connected to pig exposure. However, if the virus spreads from humans to pigs, the human population may be at increased risk because of the potential reservoir represented by the swine population. In addition, viral reassortment may be possible in the swine host, which has been previously recognized as a mixing vessel for avian and human influenza viruses. In order to investigate the potential outbreak of the new pandemic virus in pigs, a real-time reverse transcriptase-polymerase chain reaction (RT-PCR) for the detection of the 2009 pandemic H1N1 RNA in clinical specimens was developed. The assay is based on TaqMan technology, and consists of two primers with a dual-labeled probe that binds selectively to the matrix protein gene of the pandemic virus. To evaluate the applicability of the test as a diagnostic tool in the screening of field specimens from swine, we retrospectively analyzed field isolates collected during diagnostic investigations in pigs and 116 samples collected during an experimental in vivo infection with two novel H1N1 isolates, A/California/04/2009 (H1N1)v and A/Mexico/4108/2009 (H1N1)v. Our unique and highly sensitive test represents a rapid and useful approach for the screening and quantitation of the novel 2009 H1N1 RNA in clinical specimens.

Last Modified: 10/19/2014
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