Método rápido y sencillo para la recogida, conservación y envío de muestras de heces
Descripción general del producto
El sistema de recogida y conservación de hisopos fecales de Norgen está diseñado para la recogida, almacenamiento a temperatura ambiente y transporte de ácidos nucleicos de muestras de heces. Los tubos del sistema de recogida y conservación de hisopos fecales contienen el conservante de heces de Norgen en formato líquido. El usuario sólo tiene que recoger la muestra con el hisopo suministrado y, a continuación, transferirlo al conservador de heces. El conservante de heces impide el crecimiento de bacterias y hongos gramnegativos y grampositivos, y también inactiva los virus, lo que permite manipular y enviar con seguridad las muestras no infecciosas resultantes. Además, el conservante de heces elimina la necesidad de procesar o congelar inmediatamente las muestras y permite enviarlas a instalaciones de análisis centralizadas a temperatura ambiente. Los componentes del conservante de heces permiten conservar las muestras a temperatura ambiente durante más de 2 años para el ADN y 7 días para el ARN. Para prolongar la estabilidad del ARN fecal en el conservante, se recomienda almacenarlo a -20°C o -70°C desde su llegada a las instalaciones de pruebas hasta la purificación del ARN.
Detalles
Supporting Data
Figure 1. Integrity of Total Stool DNA and RNA Isolated from Preserved Stool Samples.
DNA and RNA were isolated from 200 µL of preserved stool from 4 different donors (lanes A to D) stored at room temperature at the indicated time points (DNA at 27 months and RNA at 7 days) using Norgen's Stool Nucleic Acid Isolation Kit (Cat# 45600). Next, 10 µL from the 75 µL eluted nucleic acid was loaded onto a gel for visual analysis. High integrity stool DNA was isolated from preserved stool samples that were stored at room temperature for 27 months, and high integrity RNA was isolated from stool samples stored for 7 days at ambient temperature. Lane M is Norgen's HighRanger 1 kb DNA Ladder (Cat. 11900).
Figure 2. Norgen's Best-In-Class Preservation.
Summary of community shifts in response to stabilizing solutions over a 14-day storage period. Bray-Curtis distance towards the reference for each patient grouped by stabilizing solution. Median and 5th-to-95th percentile ranges are shown for both interaliquot and interpatient variability (adapted from Plauzolles et al, 2022). Norgen is the most similar to frozen stool samples, over 11 other sample collection and storage methods.
Figure 3. Monitoring the DNA stability of Adenovirus (Ad5subE1) Spiked into Preserved Stool Samples using TaqMan Real-Time PCR Assay.
Preserved stool samples were spiked with 6 x 109 pfu/mL of Adenovirus, and 200 µL samples were removed at the indicated time points. The stool DNA was then isolated using Norgen's Stool DNA Isolation Kit (Cat# 27600). Next, 2 µL of DNA template from each 50 µL of elution was used in a 20 µL PCR reaction to detect the Adenovirus DNA. The stool preservative was shown to stabilize Adenovirus DNA up to 21 weeks at ambient temperature, indicating that Norgen's Stool Nucleic Acid Collection and Preservation Tubes provide a safe and convenient sample transportation system.
Figure 4. Hierarchical Clustering of Stool Samples at Phylum Level Under Different Storage Conditions.
The microbial composition from the stool samples preserved in Norgen’s Stool Nucleic Acid Collection and Preservation Tubes (Cat. 45630, 45660) did not differ significantly to that of controls (fresh stool from Day 0 at RT) over a 2 day period while non-preserved stool samples showed a distance in the dendrogram from the original stool profile.
Figure 5. Difference In Top 5 Abundant Microorganisms at Phylum Level Compared to Fresh Stool (Day 0).
Abundance of Firmicuties, Bacteroidetes and proteobacteria from the preserved stool sample was higher than non-preserved (NP) to the fresh stool after 2 days storage at RT, indicating that the originality of microorganism profile from the preserved are much closer to the fresh stool sample (Day 0).
Figure 6. Relative Abundance of Bacterial Genera Between Preserved And Non-preserved Stool Sample Stored at RT For 2 Days.
Preserved stool sample using Norgen’s Stool Nucleic Acid Collection and Preservation Tubes (Cat. 45630, 45660) showed closed relativeness to the fresh stool sample (Day 0).
Figure 7. Correlation between duplicate bacterial metagenomic reads.
Correlation between duplicate bacterial metagenomic reads using (A) Fresh stool (day 0) and preserved stool at RT for 2 days and (B) Fresh stool (day 0) and non-preserved stool at RT for 2 days. A greater R2 was observed for Fresh stool (day 0) and preserved stool, suggestive of higher consistency between fresh stool sample and preserved stool sample
Datos de respaldo
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Especificaciones del kit
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| Estabilidad de los ácidos nucleicos fecales a temperatura ambiente |
2 años para el ADN, hasta 7 días para el ARN*
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* La estabilidad del ARN variará en función de las muestras
Condiciones de almacenamiento y estabilidad del producto
El sistema de recogida y conservación de hisopos fecales (formato a granel) puede almacenarse a temperatura ambiente durante un máximo de 2 años sin que disminuya el rendimiento del kit. La fecha de caducidad de los hisopos está escrita en su envoltorio.