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Establishing Methods to Monitor Influenza (A)H5N1 Virus in Dairy Cattle Milk, Massachusetts, USA
Elyse Stachler

, Andreas Gnirke, Kyle McMahon, Michael Gomez, Liam Stenson, Charelisse Guevara-Reyes, Hannah Knoll, Toni Hill, Sellers Hill, Katelyn S. Messer, Jon Arizti-Sanz, Fatinah Albeez, Elizabeth Curtis, Pedram Samani, Natalia Wewior, David H. O’Connor, William Vuyk, Sophia E. Khoury, Matthew K. Schnizlein, Nicole C. Rockey, Zachariah Broemmel, Michael Mina, Lawrence C. Madoff, Shirlee Wohl, Lorraine O’Connor, Catherine M. Brown, Al Ozonoff, Daniel J. Park, Bronwyn L. MacInnis
1, and Pardis C. Sabeti
1
Author affiliation: Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA (E. Stachler, A. Gnirke, K. McMahon, M. Gomez, L. Stenson, C. Guevara-Reyes, H. Knoll, T. Hill, S. Hill, K.S. Messer, J. Arizti-Sanz, F. Albeez, E. Curtis, P. Samani, N. Wewior, A. Ozonoff, D.J. Park, B.L. MacInnis, P.C. Sabeti); University of Puerto Rico-Rio Piedras, San Juan, Puerto Rico, USA (C. Guevara-Reyes); Harvard University, Cambridge (P. Samani, B.L. MacInnis, P.C. Sabeti); University College London, London, UK (P. Samani); University of Wisconsin-Madison, Madison, Wisconsin, USA (D.H. O’Connor, W. Vuyk); The University of Texas at Austin, Austin, Texas, USA (S.E. Khoury); Michigan State University, East Lansing, Michigan, USA (M.K. Schnizlein); Duke University, Durham, North Carolina, USA (N.C. Rockey, Z. Broemmel); Immune Observatory, Boston, Massachusetts, USA (M. Mina); University of Massachusetts Chan Medical School, Worcester, Massachusetts, USA (L.C. Madoff); Massachusetts Department of Public Health, Boston (L.C. Madoff, S. Wohl, C.M. Brown); Brigham and Women’s Hospital, Boston (S. Wohl); Massachusetts Department of Agricultural Resources, Boston (L. O’Connor); Boston Children’s Hospital, Boston (A. Ozonoff); Harvard Medical School, Boston (A. Ozonoff); Howard Hughes Medical Institute, Chevy Chase, Maryland, USA (P.C. Sabeti)
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Figure 2

Figure 2. Digital PCR detection of synthetic nucleic acid (top) and RNaseP Bovine (bottom) in study of methods to monitor influenza A(H5N1) virus in dairy cattle milk, Massachusetts, USA. For direct extraction, we extracted 200 μL of milk spiked with serial dilutions of H5N1 synthetic gene fragments. For precentrifugation, we centrifuged samples for 12,000 × g for 10 minutes after spike-in, after which we extracted 200 μL. Extractions were performed using the MagMAX CORE extraction kit (Thermo Fisher Scientific, https://www.thermofisher.com).
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Page created: March 03, 2025
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