Complete Collection 2014 Price Comparison

Abstract Background Effective means are needed to efficiently collect fecal samples for microbiome analysis in large-scale epidemiological studies. Using twenty-four fecal aliquots prepared from three healthy individuals, we compared the following four fecal sample collection methods for assessment of human gut microbiome: The 24 samples were assayed by 16S rRNA gene sequencing to compare overall microbiome structure and taxon distributions according to collection method.

Results Storing fecal occult blood test card samples at room temperature for three days did not affect total DNA purity and relative 16S rRNA bacterial gene contents, compared with fresh frozen collection. Conclusions Our findings suggest that low-cost fecal occult blood test card collection may be a feasible means of sample collection for fecal microbiome assessment in large-scale population-based studies.

Background The human bowel hosts trillions of gut microbial cells, the gut microbiome[ 1 ]. Although case—control investigation points to a potential role of the gut microbiome in colorectal cancer[ 2 ], large-scale prospective study of this association has been impeded by the lack of validated fecal sample collection methods suitable for large-scale studies. Our interest was in development of a fecal sample collection method that is accurate, while also being cost-efficient and easy for the study participant to use.

Because fecal collections may take place outside of research clinics, we also wished to develop a fecal collection approach which would not require immediate sample processing. Finally, we were interested in evaluating the utility of collection in RNAlater, because this RNA-stabilizing agent has been shown to be suitable for samples dedicated for DNA amplification[ 5 , 6 ].

Our main goal was to assess the effect the different storage conditions have on gut microbiome diversity parameters including overall diversity and specific taxon abundances because different methods might differentially impact bacterial overgrowth or DNA degradation that could lead to biased assessment of these microbial parameters.

In Method 2 room temperature fecal samples were placed in an Eppendorf tube and left at room temperature. All samples were stored for three days in their respective method.

We chose three days to mimic the conditions of samples being collected at home and returned to us by mail. All study participants provided informed consent. Chimeric sequences were identified by ChimeraSlayer[ 10 ] and removed. A phylogenetic tree was constructed by applying the FastTree method[ 12 ] to the representative sequences.

Rarefactions of 10 to 8, [minimum-maximum sequence depth] randomly selected sequences from each sample were used to calculate the Shannon index, a measure of within sample diversity, and to generate rarefaction plots. Pairwise comparisons of Shannon indices by subject and storage condition were obtained by Monte Carlo permutation. All p-values were adjusted by Bonferroni correction. To measure the diversity among subjects or storage conditions, a single rarefaction was performed at a sequencing depth of so that all samples were included in analyses.

Principal coordinates were computed for the unweighted distance matrices and used to generate Principal Coordinate Analysis plots PCoA. The non-parametric method, adonis[ 14 ], was used to identify significant differences in phylogenetic distance variation by subjects and by storage condition. A two-sample t-test was used to test for differences between the within and between group variances, with p-values adjusted by Bonferroni correction.

Relative abundances of the three major phyla Bacteroidetes, Firmicutes, Actinobacteria were compared for the four methods, using the Mann—Whitney-Wilcoxon test, and compared by subject, using the Kruskal-Wallis test SAS, version 9. Results DNA from 24 fecal aliquots was successfully extracted and amplified.

From the initial , microbial 16S rRNA sequences, , sequence reads passed filtering criteria. A , B The first two principal coordinates were grouped by subject 1 [red], 2 [blue], 3 [orange] A or collection method card [green], room temperature [blue], RNAlater [orange], frozen [red] B.

Adonis was used to test for significant differences in the variation in distances across subjects or collection methods using QIIME. Full size image Relative abundances of gut microbial taxa were not statistically different for any of the three test methods, when compared to relative abundances from frozen samples. The average relative abundances for the three major phyla were Firmicutes Figure 3 Relative abundances of phyla by subject and by collection method.

Full size image Discussion We found no evidence of significant differences in gut microbial community composition and taxon distributions for storage at room temperature on a fecal occult blood test card or in an Eppendorf tube compared to immediately frozen samples. Not surprisingly, overall microbial diversity varied by subject. Although the effect of collection container has not been previously assessed, our general observation that inter-individual differences in bacterial composition were greater than the differences by collection method is consistent with findings from previous studies.

RNAlater is used to stabilize and protect RNA from degradation in tissue during long term storage and has been shown to also be suitable for DNA preservation[ 21 ]. However, we observed that fecal samples were very hard to disperse evenly in RNAlater during processing and that DNA purity was lower. Low-quality DNA can interfere with downstream applications including PCR amplification[ 22 ], a possible reason for the trend toward reduced Shannon indices.

While the first study showed that total DNA yields from RNAlater samples were higher compared to refrigeration storage and liquid nitrogen freezing, the impact on Shannon indices was not described[ 5 ]. RNAlater storage increases the potential utility of stored fecal samples, so further study is warranted to determine the conditions of collection for which this reagent is suitable. Although our study showed no differences in microbiome composition between card collection with room temperature storage and collection in Eppendorf tube with immediate freezing, we recognize that a larger series of samples may have identified some differences not found here.

Also, our subjects were healthy and the collected samples may not have captured the full range of stool conditions that might be expected if subjects were ill. These considerations may be important in carrying out stool collection in different study settings. Our findings support the use of fecal occult blood test card collections for microbiome assessment of fecal samples. These cards are commercially available and inexpensive. The small size and flat shape also makes the card easier to include in packages to be sent to participants, compared to bulkier Eppendorf tubes.

Study subjects can easily collect samples on the cards. Because the cards are widely used in colorectal cancer screening[ 23 ], potential participants might also be more accepting of collecting samples in this way. However, we found no evidence of a significant difference in gut microbiome in fecal samples collected by this method.

Findings that results were unaffected by three-day storage at room temperature of the collection cards or Eppendorf tubes suggests that participant home-collection and mailing of these samples is suitable for epidemiological studies. Conclusions Our findings suggest that fecal collection on a fecal occult blood test card or in an Eppendorf tube and storage for three days at room temperature does not substantially influence the assessment of gut microbiome.

Because of the low-cost and simplicity of use, fecal occult blood test card collection may be a feasible method for large-scale population-based studies.

Downton Abbey DVDs

llll➤ Stella DVDs – Cheapest prices with our free Stella box set price comparison ⚖. Stella Series 6 [DVD] · Stella: The Complete Collection [DVD] · Stella Box Set Stella - Series 4 + Christmas Special [DVD] [] · Stella - Series 3. Buy Mount & Blade Collection CD KEY Compare Prices. Activate the CD KEY on your Steam client to download Mount & Blade Collection. Complete: Complete Edition. .. Taavi Pillesaar says on August 08, , Log in to Reply. 16th November See full specifications, expert reviews, user ratings, and more. Compare Samsung Galaxy S5 prices before buying online. Samsung Galaxy S5 smartphone was launched in April The phone comes . This is small but big collection of technology that shake the techno world One world one. May 8, - Producer price index for hospital inpatient care, by payer, . Average price full knee replacement in large employer plans, -.

Complete Collection 2014 Price Comparison

Related Links Here: Cheap Microsoft SQL Server 2014 Enterprise Software | The Best Deals On Spectrasonics Omnisphere Software

Tags:Paragon, Painter, Kaspersky

Copyrights ©2019 All rights reserved. Site Map