Toothbrushes as a Source of DNA for Gender and Human Identification—A Systematic Review
Abstract
:1. Introduction
2. Materials and Methods
- (1)
- Original research articles that have assessed and quantified DNA and/or genes from toothbrush samples;
- (2)
- Original studies that have compared the quantity or quality of DNA from toothbrushes at various time intervals;
- (3)
- Original studies that have compared the quantity or quality of DNA from different methods of DNA extraction;
- (4)
- Study design was not a case report, editorial, letter to the editor, or review;
- (5)
- The report was published in the English language.
3. Results
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Acknowledgments
Conflicts of Interest
References
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S. No. | Author Name/Year/Country | Study Design | Sample Size | Study Groups | Type of Toothbrushes | Duration of Use of Toothbrushes | Method of DNA Extraction | Gene(s) Assessed with Amplification Method | Capillary Electrophoresis Instrument | Results | Inference/Statistical Significance |
---|---|---|---|---|---|---|---|---|---|---|---|
1. | Alfadaly et al./2016/Saudi Arabia | Observational cross-sectional study | Total number of patients: 25; Total number of toothbrushes: 104; (25 uncovered siwaks, 4 covered siwaks, and 25 toothbrushes) | Group I (4 months, 4 volunteers); Group II (3 months, 5 volunteers); Group III (2 months, 4 volunteers); Group IV (1 month, 5 volunteers); Group V (1 week, 3 volunteers); Group VI (the same day, 4 volunteers, and covered siwak was added); Group VII (reference samples); Group VIII (positive and negative control samples) | Toothbrush, uncovered siwak, and covered siwak | 1 week, 1 month, 2 months, 3 months, and 4 months | Promega kit | DNA profiling: AmpFLSTR Identifiler PCR Amplification Kit | 3130XL Genetic Analyzer and GeneMapper software | DNA from siwak samples (18.96 ± 16.15) was higher than toothbrush samples (1.76 ± 1.07). 1/25 samples in siwak only partial DNA profiling could be done. In toothbrush group 8/25 profiling could be done and 3/25 partial profiling was done. Unit of DNA measurement was ng/μL | Significant (p < 0.001%) |
2. | Reddy et al./2011/India | Observational cross-sectional study | 30 samples, 30 patients (each used a sample for a week). Equal gender distribution in each group | Group 1: 10 samples immediately processed; Group II: 10 samples processed after 1 month; Group III: 10 samples processed after 2 months | Toothbrush brand name not mentioned | Immediately, 1 month, and 2 months | Real Genomics YGB 100 (Real Biotech Corporation, Taiwan) DNA extraction kit | SRY gene for gender identification using real-time PCR and Taq PCR Master Mix (Qiagen, India) | Real Plex Master Cycler (Eppendorf, Japan) | Genetic material was preset in low quantity in most of the samples. Gender identification: All males were identified correctly; out of 15 females, 4 were wrongly identified. Unit of DNA measurement was ng/μL | Sensitivity of SRY gene was 100%. Specificity of SRY gene in gender was 73.33% |
3. | Riemer et al./2012/Canada | Observational cross-sectional study | N = 55 (25 males and 30 females) | Group I: 21 individuals used their toothbrush for 1 month; Group II: 22 individuals used their toothbrush for 3 months; Group II: 12 individuals gave their currently used toothbrush for analysis. Negative control: 2 unused toothbrushes | Toothbrush brand name not mentioned | Current used toothbrush, 1 month, and 3 months | Partial toothbrush head sampling technique phenol-chloroform method, and profiles were obtained using AmpFISTR Profiler Plus (Applied Biosystems, Foster City, CA) | DNA profile compared with reference standard AmpFISTR Profiler Plus PCR Identification Kit (Applied Biosystems, Foster City, CA) | No data available | DNA yield: 600 ng of DNA per toothbrush. DNA profiling complete in 51 and partial in 4 samples. Unit of DNA measurement was ng/g | No significant difference in the quantity and quality of DNA obtained from 1 month, 3 months, and random period used toothbrushes |
4. | Tanaka et al./2000/Japan | Observational cross-sectional study | 10 toothbrushes from 10 individuals | Case toothbrush samples (among 10 samples, 3 were actual cases: 1 drowned patient and 2 from murder case). Control: blood samples from 8 patients (blood samples from the heart of both the drowning and homicide victims were obtained at autopsy) | Used toothbrush | 3 months to 1 year. Among 10 samples, 1 was stored for 6 months before analysis | Whole toothbrush head was taken for DNA extraction by phenolchloroform extraction and ethanol precipitation. Quantification by fluorometry | Six loci (DQA1, LDLR, GYPA, HBGG, D7S8, and GC); Nine STR loci (D3S1358, vWA, FGA, TH01, TPOX, CSF1PO, D5S818, D13S317, and D7S820). AmpFISTR Profiler Plus PCR Amplification Kit (Applied Biosystems, Foster City, CA) | 310 Genetic Analyzer (Applied Biosystems, Foster City, CA) | 9 toothbrushes: 10 to 430 ng; 10 toothbrushes: 0.5 ng mL. All loci were types in all samples despite low DNA yield. Unit of DNA measurement was ng/μL | All the test samples were typed at all loci. (No statistical analysis was performed) |
5. | Bandhaya et al./2007/Thailand | Observational cross-sectional study | Total samples obtained from 4 individuals who used toothbrushes for 1, 7, 14, or 30 days | Group 1: toothbrush used for 1 day; Group 2: toothbrush used for 7 days; Group 3: toothbrush used for 14 days; Group 4: toothbrush used for 30 days. Each group had the following subgroups: Sub group a1: 5 bristle bundles DNA extraction done by Chelex1-100; Sub group a2: 5 bristle bundles DNA extraction done by QIAamp DNA Mini Kit; Subgroup b1: 10 bristle bundles DNA extraction done by Chelex1-100; Sub group b2: 5 bristle bundles DNA extraction done by QIAamp DNA Mini Kit | Used toothbrushes | 1, 7, 14, and 30 days | Chelex1-100 or QIAamp DNA Mini Kit | STR (GeneAmp PCR System 9700) (Applied Biosystems) | ABI PRISM 3100 Genetic Analyzer and GeneMapper ID Software (Applied Biosystems) | DNA from QIAamp DNA Mini Kit > DNA from Chelex1-100 kit. Complete profile could be typed from QIAamp DNA Mini Kit but not in Chelex1-100 kit. Comparing 5 and 10 bristle bundles, STR was complete in all samples of Mini Kit from 5 bristle bundles and was complete in 30- and 14-day samples of 10 bristle bundles of Mini Kit. In the Chelex1-100 kit, none was completely typed. Unit of DNA measurement was ng/μL | Statistical analysis was not done. DNA Mini Kit was a better method for DNA extraction and 5 bristle bundles are better suited for DNA extraction from toothbrushes |
S. No. | Checklist | Alfadaly et al./2016/Saudi Arabia | Reddy et al./2011/India | Riemer et al./2012/Canada | Tanaka et al./2000/Japan | Bandhaya et al./2007/Thailand |
---|---|---|---|---|---|---|
1 | Were the aims/objectives of the study clear? | Yes | Yes | Yes | Yes | Yes |
2 | Was the study design appropriate for the stated aim(s)? | Yes | Yes | Yes | Yes | Yes |
3 | Was the sample size justified? | No | No | No | No | No |
4 | Was the target/reference population clearly defined? (Is it clear who the research was about?) | Yes | Yes | Yes | Yes | Yes |
5 | Was the sample frame taken from an appropriate population base so that it closely represented the target/reference population under investigation? | Unclear | Unclear | Unclear | Unclear | Unclear |
6 | Was the selection process likely to select subjects/participants that were representative of the target/reference population under investigation? | No | No | No | No | No |
7 | Were measures undertaken to address and categorize non-responders? | Yes | Yes | Yes | Yes | Yes |
8 | Were the risk factor and outcome variables measured appropriate to the aims of the study? | Not applicable | Not applicable | Not applicable | Not applicable | Not applicable |
9 | Were the risk factor and outcome variables measured correctly using instruments/measurements that had been trialed, piloted, or published previously? | Not applicable | Not applicable | Not applicable | Not applicable | Not applicable |
10 | Is it clear what was used to determined statistical significance and/or precision estimates? (e.g., p-values, CIs) | Yes | Yes | Yes | No | No |
11 | Were the methods (including statistical methods) sufficiently described to enable them to be repeated? | No (statistical analysis was not described) | No (statistical analysis was not described) | No (statistical analysis was not described) | No (statistical analysis was not described) | No (statistical analysis was not described) |
12 | Were the basic data adequately described? | Yes | Yes | Yes | Yes | Yes |
13 | Does the response rate raise concerns about non-response bias? | Yes | Yes | Yes | Yes | Yes |
14 | If appropriate, was information about non-responders described? | Yes | Yes | Yes | Yes | Yes |
15 | Were the results internally consistent? | Yes | Yes | Yes | Yes | Yes |
16 | Were the results for the analyses described in the methods, presented? | Yes | Yes | Yes | Yes | Yes |
17 | Overall risk of bias | Low | Low | Low | Moderate | Moderate |
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Sujatha, G.; Priya, V.V.; Dubey, A.; Mujoo, S.; Sulimany, A.M.; Omar Tawhari, A.M.; Mokli, L.K.; Mohana, A.J.; Varadarajan, S.; Balaji, T.M.; et al. Toothbrushes as a Source of DNA for Gender and Human Identification—A Systematic Review. Int. J. Environ. Res. Public Health 2021, 18, 11182. https://doi.org/10.3390/ijerph182111182
Sujatha G, Priya VV, Dubey A, Mujoo S, Sulimany AM, Omar Tawhari AM, Mokli LK, Mohana AJ, Varadarajan S, Balaji TM, et al. Toothbrushes as a Source of DNA for Gender and Human Identification—A Systematic Review. International Journal of Environmental Research and Public Health. 2021; 18(21):11182. https://doi.org/10.3390/ijerph182111182
Chicago/Turabian StyleSujatha, Govindarajan, Veeraraghavan Vishnu Priya, Alok Dubey, Sheetal Mujoo, Ayman M. Sulimany, Ali Mohammed Omar Tawhari, Lujain Khalawi Mokli, Arwa Jaber Mohana, Saranya Varadarajan, Thodur Madapusi Balaji, and et al. 2021. "Toothbrushes as a Source of DNA for Gender and Human Identification—A Systematic Review" International Journal of Environmental Research and Public Health 18, no. 21: 11182. https://doi.org/10.3390/ijerph182111182