Lab Matters Spring 2026 | Page 13

INDUSTRY MATTERS

By Kaylinnette Pinet, PhD, development scientist II, Applications & Product Development, New England Biolabs and Betsy Young, PhD, senior product marketing manager, Next Generation Sequencing, New England Biolabs

Highly pathogenic avian influenza( HPAI) A virus continues to evolve across animal reservoirs. While zoonotic transmission to humans in the United States has been limited, recent epidemiological trends in Cambodia underscore how quickly zoonotic risks can escalate. After nearly a decade without human cases, Cambodia has seen a resurgence of H5N1 infections, highlighting dynamic viral evolution across poultry and humans. According to the World Health Organization( WHO), Cambodia saw a total of 27 H5N1 cases from 2023 to July 2025, with 12 associated deaths. Across 25 countries, during the same time frame, WHO reported an H5N1 fatality ratio of 48 %, reinforcing the need for vigilant surveillance.

Over the years, the international movement of infected animal products or contaminated materials has dramatically increased due to trade and shipping route expansions. Protecting the international food supply, for US consumers and others, will involve improving rapid agricultural sample sequencing capacity. This is particularly relevant to poultry and dairy farmers and processors. inhibitor-tolerant extraction with an easy, one-step RT-PCR and indexed influenza A primers for whole-genome amplification and library preparation. Ultimately, this workflow transforms these samples into data in hours rather than days.

• Monarch ® Mag Viral DNA / RNA Extraction Kit( NEB # T4010) This magnetic bead-based system is optimized for complex matrices. It efficiently extracts and purifies nucleic acids from fats, proteins, and PCR inhibitors while preserving viral RNA integrity. The result is highquality RNA suitable for sensitive RT-PCR, qPCR, LAMP and sequencing applications.

• LunaScript ® Multiplex One-Step RT-PCR Kit( NEB # E1555) Following extraction, LunaScript enables robust reverse transcription and amplification of Influenza A gRNA in a single step. Its optimized chemistry supports multiplexed amplification, reducing hands-on time and improving sensitivity for low-abundance targets.

• NEBNext ® Flu A Integrated Indexing Primer Module( NEB # E3436) This module enables amplification of all eight influenza A genome segments and incorporates indexing in a single step. This design reduces workflow complexity, minimizes errors and promotes comprehensive sequencing with Oxford Nanopore Technologies ® Ligation Sequencing Kit V14 / XLV14( or Illumina ® sequencing).

Finding Flu A, Closer to Home

The US has most recently seen HPAI transmission to humans via milk products, raising concerns about consumer safety, warranting additional safety screening. However, milk is a notoriously difficult matrix for viral detection due to its constituent fats, proteins and enzymatic inhibitors that can suppress extraction efficiency and impede downstream assays, leading

continued on page 17

A new correspondence in the New England Journal of Medicine from Institut Pasteur du Cambodge demonstrated that rapid and routine sequencing of poultry swabs( e. g., cloacal, oropharyngeal) can quickly detect influenza A genome reassortment events and identify markers linked to increased virulence and transmissibility. These types of data could help guide interventions before viruses break through into wider supply chains, putting the US food supply and human health in jeopardy.

A Purpose-Built Workflow for Influenza A Surveillance

The workflow( Figure 1) was inspired by the need for prompt agricultural sample surveillance and is supported by reagents from New England Biolabs. It pairs

New England Biolabs is an APHL Gold Level Sustaining Member.

Figure 1. Finding Flu A, Faster. The complete NEB workflow for influenza A identification begins with RNA extraction and purification, followed closely by reverse transcription and amplification. With optional library prep for influenza A sequencing, these solutions are fast and reliable.