GT-Fast Prep Influenza A Library Kit for Illumina®
Description
(Cat# 101120)
The GT-Fast Prep Library Kit for Illumina is a molecular reagent kit containing all primers and reagents for the amplification and downstream library preparation of influenza A gRNA extracts from wastewater matrices. GT Molecular’s high-performance workflow, in combination with the proprietary GT Molecular primer set, provides researchers with and ≥85% coverage of the hemagglutinin gene. This assay is designed for high confidence subclade resolution and base level mutation tracking while monitoring influenza A viral outbreaks.
This kit is optimized for use with wastewater extracts known to be positive for the presence of influenza A, as determined by digital or Real-Time PCR. The GT-Fast Prep Influenza A Library Kit for Illumina was developed using historical data on influenza A from the past 50 years. Although this kit can detect novel variants, there may be some loss of uniformity or coverage if variations, particularly insertions and deletions, are present within underlying primer sequences. Please contact us for the latest data regarding the use of this product with the season’s circulating strains.
Note: A positive control is not included in this kit but is available to purchase separately. The FluA H3N2 Positive Control (Cat# 101011) is a synthetic RNA corresponding to Influenza A H3N2 which can be used for dPCR, RT-PCR, or NGS assays targeting Influenza A (H3N2). It is intended for use as a semi-quantitative process and performance control in NGS, dPCR, or RT-PCR assays targeting Influenza A (H3N2). For NGS assays, the control can also be used to assess assay accuracy and error rates by aligning the sequenced data against the provided reference genome.
GT molecular is proud to provide the first Flu A sequencing kit validated and optimized for wastewater.
Requirements
This kit has been validated for the preparation of NGS libraries from wastewater samples extracted utilizing the Promega wastewater workflow (Maxwell® RSC Enviro TNA kit, (Cat#AS1831)) or CERES wastewater workflow (CERES Nanotrap® Microbiome A Particles (Cat#44202-30), CERES Nanotrap® Enhancement Reagent 1 (Cat#10111-30)) and Thermo Fisher Scientific (MagMAX™ Wastewater Ultra Kit (Cat#A52606)). Please contact us for extraction workflow recommendations. The kit has been designed with a limit of detection of 5 copies of influenza A per µL of processed nucleic acid wastewater extract, as measured by dPCR. Samples below this concentration are not recommended, but users can choose to proceed at their own discretion. Low concentration samples will generate lower coverage, higher off-target reads, and will require deeper sequencing. We recommend using the GT-Digital Flu;SC2;RSV Wastewater Surveillance Panel (Cat# 100735 or 100740) to identify and quantify positive influenza A from wastewater extracts. GT Molecular does NOT recommend the use of the GT-Fast Prep Influenza A Library Kit for Illumina for the detection or quantitation of influenza A: for quantification assay information and ordering, please contact us.
Technical Note
Uneven coverage across the eight influenza A genome segments is a biologically deterministic outcome of (1) the intrinsic molecular architecture of influenza virions, (2) the physio-chemical environment of wastewater, and (3) segment–specific differences in RNA stability, structure, and packaging. Multiple mechanisms may affect the segment coverage before any extraction, reverse transcription (RT), or sequencing step, establishing a biologically imposed ceiling on viral genomic completeness. It is important to note that technical optimization can reduce but never eliminate dropouts caused by intrinsic biological processes, specific differences in RNA stability, structure, and packaging. To overcome these constraints, this kit uses a highly sensitive, subtype-specific sequencing approach tailored to wastewater. Instead of relying on universal primers, the kit uses targeted primer sets designed explicitly for the human seasonal influenza subtypes H1N1 and H3N2. This design ensures the kit captures human–relevant sequences while avoiding detection of avian–origin strains like H5N1. For more details on the technical limitations of sequencing influenza A genomic RNA from wastewater, please contact support@gtmolecular.com
Positive Control
A positive control is not included in this kit but is available to purchase separately. The FluA H3N2 Positive Control (Cat# 101011) is a synthetic RNA corresponding to Influenza A H3N2 which can be used for dPCR, RT-PCR, or NGS assays targeting Influenza A (H3N2). It is intended for use as a semi-quantitative process and performance control in NGS, dPCR, or RT-PCR assays targeting Influenza A (H3N2). For NGS assays, the control can also be used to assess assay accuracy and error rates by aligning the sequenced data against the provided reference genome.