| Technology | Silica membrane technology |
| Format | Single-spin format |
| Sample amount | 200 μl serum 200 μl cell-free biological fluids, e.g., milk 5–10 mg tissue 100 μl animal blood 100 mg feces/stool 1 dry or wet swab |
| Fragment size | Approx. 100 bp–50 kb |
| Elution column volume | 40–100 μl |
| Processing time | 20–40 min |
Product Overview
Extraction of viral DNA/RNA and bacterial DNA from veterinary samples with NucleoSpin VET
Modern veterinary diagnostic laboratories must be able to collect and subsequently analyze hundreds of samples fully automatically every day—especially in times of disease outbreaks. Despite high levels of automation, many samples still require manual nucleic acid purification. This is particularly the case when there is a low sample load, when difficult and inhomogeneous samples (e.g., feces) must be analyzed, or when new methods are to be established in a veterinary laboratory. NucleoSpin VET is a single-spin solution for the isolation of viral RNA/DNA and bacterial DNA from various veterinary sample materials, including serum, plasma, animal whole blood, tissue, feces, swabs, and other cell-free biological samples.
- One standard protocol for all specimen matrices after sample pretreatment
- Removes inhibitors and contaminants, even when working with difficult sample matrices (e.g., feces or fatty raw milk)
- Yields eluates that can be directly used for all common downstream analysis methods, including RT-qPCR and NGS
- Includes FastTrack protocol for rapid, routine testing that is focused on speed and ease of use combined with maximum sensitivity
At-a-glance
Protocols
NucleoSpin VET enables efficient purification of viral RNA and DNA, as well as bacterial DNA, from a broad range of veterinary-relevant sample types. All samples can be purified following the same standard protocol after sample lysis. The sample lysis pretreatment depends on starting material and target pathogen.
Figure 1. Standard NucleoSpin VET nucleic acid purification workflow.
NucleoSpin VET also includes a FastTrack protocol for rapid, routine testing of low-complexity sample matrices (e.g., plasma, swab solutions; see user manual for the protocol). The FastTrack protocol has been optimized in terms of pipetting steps, standardized volumes, and centrifuge setting changes, allowing fast and simple sample extraction. We recommend that users start with the standard protocol first and validate the use of the FastTrack protocol for their specific sample material.
Figure 2. Comparison of NucleoSpin VET standard and FastTrack protocols. Canine distemper virus (CDV) (Panel A) or porcine enterovirus (PEV) (Panel B) were spiked into bovine plasma or saliva samples. Then, nucleic acids were isolated from spiked samples using the standard or FastTrack NucleoSpin VET protocols, and recovery of CDV and PEV were assessed via RT-qPCR. Ct values were comparable for the standard and FastTrack protocols.
Example data
Figure 3. Reliable viral DNA and RNA recovery from bovine blood with NucleoSpin VET. The NucleoSpin VET kit was used for viral DNA and RNA isolation from bovine blood. Thawed bovine blood was spiked with lumpy skin disease virus (LSDV), bovine coronavirus (BoCoV), and foot-and-mouth disease virus (FMDV) at various dilution levels. Virus-spiked blood samples were purified using NucleoSpin VET and eluates were analyzed via RT-qPCR using AgPath-IDTM One-Step RT-PCR reagents (ThermoFisher Scientific). The NucleoSpin VET kit demonstrated consistent and reliable viral RNA and DNA recovery from high and low viral titer samples (FMDV titer in 1:10,000 dilution was below detection limit). Data kindly provided by Hanna Keck and Michael Eschbaumer, Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health of Germany.
Figure 4. Market-leading performance compared to competitor products. Extractions from cloacal swab solutions, chicken liver tissue, bovine blood, and bovine plasma samples were spiked with MS2 viral RNA. Nucleic acid purifications were performed using NucleoSpin VET or using an extraction kit from Competitor I. Following nucleic acid extraction, RT-qPCR analysis was performed using the Bioline SensiFAST SYBR Lo-ROX One-Step Kit to determine the viral titer load in the different sample matrices. Extraction with NucleoSpin VET resulted in higher viral RNA recovery compared to Competitor I’s kit.
Figure 5. Comparison of manual and automated extraction methods in detection of foot-and-mouth disease virus in bovine milk and skin lesions. Nucleic acids were isolated from foot-and-mouth disease virus (FMDV)-positive bovine milk and skin lesions using NucleoSpin VET (manual single spin purification) or NucleoMag VET (magnetic bead-based automated extraction). For the NucleoMag VET, 100 μl sample input was used. Eluates were subsequently analyzed via RT-qPCR using AgPath-IDTM One-Step RT-PCR reagents (ThermoFisher Scientific). Ct values for each sample extracted using NucleoSpin VET were comparable to samples extracted using NucleoMag VET. Data kindly provided by Hanna Keck and Michael Eschbaumer, Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health of Germany.
Figure 6. Comparison of manual and automated extraction methods for high to low viral titer samples. Varying dilutions of MS2 viral RNA were spiked into bovine plasma to simulate low, medium, and high viral titer samples. Nucleic acid purifications were performed using NucleoSpin VET (manual single-spin purification) or NucleoMag VET (magnetic bead-based automated extraction) and analyzed via RT-qPCR using the Bioline SensiFAST SYBR Lo-ROX One-Step Kit. Ct values for low, medium, and high viral titer samples extracted using NucleoSpin VET were comparable to those samples extracted using NucleoMag VET.
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