| Literature DB >> 24288659 |
Alex M Garvin1, Ralf Holzinger, Florian Berner, Walter Krebs, Bernhard Hostettler, Elges Lardi, Christian Hertli, Roy Quartermaine, Christoph Stamm.
Abstract
Biological samples are vulnerable to degradation from the time they are collected until they are analysed at the laboratory. Biological contaminants, such as bacteria, fungi, and enzymes, as well as environmental factors, such as sunlight, heat, and humidity, can increase the rate of DNA degradation. Currently, DNA samples are normally dried or frozen to limit their degradation prior to their arrival at the laboratory. In this study, the effect of the sample drying rate on DNA preservation was investigated, as well as a comparison between drying and freezing methods. The drying performances of two commercially available DNA collection tools (swab and drying tube) with different drying rates were evaluated. The swabs were used to collect human saliva, placed into the drying tubes, and stored in a controlled environment at 25°C and 60% relative humidity, or frozen at -20°C, for 2 weeks. Swabs that were stored in fast sample drying tubes yielded 95% recoverable DNA, whereas swabs stored in tubes with slower sample drying rates yielded only 12% recoverable DNA; saliva stored in a microtube at -20°C was used as a control. Thus, DNA sampling tools that offer rapid drying can significantly improve the preservation of DNA collected on a swab, increasing the quantity of DNA available for subsequent analysis.Entities:
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Year: 2013 PMID: 24288659 PMCID: PMC3830855 DOI: 10.1155/2013/105797
Source DB: PubMed Journal: Biomed Res Int Impact factor: 3.411
Figure 1forensiX evidence collection tube, disassembled, with cotton swab and the SafeDry desiccant insert with a serrated inner surface.
Figure 2Sarstedt Forensic swab with disassembled ventilation membrane and viscose swab.
Summary of saliva-loaded tubes and storage conditions.
| Sample no. | Type of storage tube | Storage condition |
|---|---|---|
| 1–3 | Microfuge tube (positive control) | Frozen (−20°C) |
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| 4–6 | forensiX evidence collection tube; cotton swab with SafeDry desiccant | Frozen (−20°C) |
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| 7–9 | Sarstedt forensic tube with ventilation membrane | Frozen (−20°C) |
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| 10–12 | forensiX evidence collection tube without SafeDry desiccant | 25 ± 2°C at 60% relative humidity |
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| 13–15 | forensiX evidence collection tube; cotton swab with SafeDry desiccant | 25 ± 2°C at 60% relative humidity |
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| 16–18 | Sarstedt forensic tube with membrane sealed using Scotch tape | 25 ± 2°C at 60% relative humidity |
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| 19–21 | Sarstedt forensic tube with ventilation membrane | 25 ± 2°C at 60% relative humidity |
Collection tubes and storage conditions for microbial growth assays.
| Sample no. | Type of storage tube | Storage conditions |
|---|---|---|
| 1–3 | forensiX evidence collection tube | 25 ± 2°C at 60% relative humidity; 14 days |
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| 4–6 | Sarstedt forensic tube | 25 ± 2°C at 60% relative humidity; 14 days |
Figure 3Drying curve for the initial 24 hours: the percentage of remaining water on swabs in collection tubes (n = 5) at 24 ± 2°C with a relative humidity of 50 ± 5%.
Figure 4Drying curve over 14 days: the percentage of remaining water on swabs in collection tubes (n = 5) at 24 ± 2°C with a relative humidity of 50 ± 5%.
DNA yields for the different storage methods.
| Samples | Description of storage of saliva sample | Average DNA yield per swab (ng) | Standard deviation of results (ng) | DNA recovery relative to saliva controls |
|---|---|---|---|---|
| 1–3 | Microfuge tube, frozen, and control | 455 | 47.4 | 100.0% |
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| 4–6 | forensiX Swabs, frozen | 328 | 75.4 | 72.0% |
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| 7–9 | Sarstedt Swabs, frozen | 246 | 20.3 | 54.0% |
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| 10–12 | forensiX Swabs, without desiccant | 22 | 4.5 | 4.8% |
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| 13–15 | forensiX Swabs with desiccant | 431 | 13.8 | 95.0% |
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| 16–18 | Sarstedt Swabs, sealed | 4 | 2.0 | 0.9% |
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| 19–21 | Sarstedt Swabs, unsealed | 56 | 3.3 | 12.3% |
Yield of saliva DNA from different swab materials (n = 3 × 2 = 6).
| Samples | Description of storage of saliva sample | Conditions | Average DNA yield per swab (ng) | Standard deviation of results (ng) |
|---|---|---|---|---|
| 1–3 | Sarstedt swab (viscose) | Air dried: 56°C, 2 hr | 331 | 84 |
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| 4–6 | forensiX swab (cotton) | Air dried: 56°C, 2 hr | 309 | 76 |
Colony numbers on SDA plates.
| Sample type | forensiX evidence collection tube stored at 25°C, 60% relative humidity over 14 days | Sarstedt swab stored at 25°C, 60% relative humidity over 14 days | ||||
|---|---|---|---|---|---|---|
| Swab no. |
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| No dilution | <1 | <1 | <1 | 5 | >250 | >250 |
| 1 : 10 dilution | <1 | <1 | <1 | <1 | 39 | 19 |
| 1 : 100 dilution | <1 | <1 | <1 | <1 | 5 | <1 |
Genus level identification of bacterial colonies obtained from Sarstedt tubes.
| Genus | Number of colonies | Percent of total |
|---|---|---|
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| 18 | 56% |
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| 7 | 22% |
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| 6 | 19% |
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| 1 | 3% |
Figure 5Temporal changes in the ATP content of P. putida during 24 hours on swabs in forensiX tubes with or without a desiccant (n = 3).