Isolation and quantification of DNA from epithelial cells obtained from acrylic removable partial denture for sex identification

Abstract

Background: This study is aimed to evaluate the use of epithelial cells adhering the acrylic removable partial denture in establishing the sex of the individual using sex-determining region of the Y chromosome (SRY) gene. With a hypothesis that prosthetic appliance can also be used as a source of DNA in forensics for sex determination. A sample of 30 partial dentures from different denture wearers was selected for this study. Informed consent was taken from the subjects and institutional ethical clearance was obtained for this study. Samples were then washed in saline and scrapings from 0.5 × 0.5 cm 2 of the tissue contacting surface were taken using a blade of fixed thickness. DNA extraction was done from these isolated epithelial cells using Real Genomics YGB 100 (Real Biotech Corporation, Taiwan). Quantification and purity assessment of the extracted DNA was done using a Nanodrop ND-1000 spectrophotometer and clean DNA was subjected to real-time PCR amplification using SRY sex-typing marker for the detection of the presence of the SRY gene. Results: Isolation of epithelial cells and extraction of DNA from all 30 acrylic removable partial dentures were possible. Average DNA yield quantified from all samples was 32.37 ng/μl with a minimum yield of 2.17 ng/μl and a maximum yield of 117.57 ng/μl. On amplification of the target sequence, sex determination was possible from all samples with 100% accuracy. Conclusion: Acrylic removable partial denture can also be used as a source of DNA in forensics for sex identification.

Molecular analysis for sex determination in forensic dentistry: a systematic review

“…The results showed that most of the studies succeeded in DNA extraction except for two studies (Srivastava et al 2017;Chowdhury et al 2018). The sex identification was accurately defined in six studies (Zapico and Ubelaker 2013;Chauhan et al 2016;Kholief et al 2017;Dutta et al 2017;Bharath et al 2019), and in four studies, not all samples were identified correctly (Reddy et al 2011;Srivastava et al 2017;Chowdhury et al 2018;Lim et al 2019). The conclusion from ten studies was that teeth, mesiodens, dental pulp, and dentin are reliable sources for DNA with resistance to forensic insult unless subjected to extreme degradation and that also toothbrushes and dental prostheses were good DNA sources.…”

“…The conclusion from ten studies was that teeth, mesiodens, dental pulp, and dentin are reliable sources for DNA with resistance to forensic insult unless subjected to extreme degradation and that also toothbrushes and dental prostheses were good DNA sources. Amelogenin (Zapico and Ubelaker 2013;Nakanishi et al 2015;Chauhan et al 2016;Srivastava et al 2017;Dutta et al 2017;Chowdhury et al 2018;Lim et al 2019), SRY (Reddy et al 2011;Kholief et al 2017;Bharath et al 2019), and DYS14 (Kholief et al 2017) were all reliable in sex identification.…”

“…These ten studies included 335 samples, and three studies mention samples originating from male and female subjects (Nakanishi et al 2015;Kholief et al 2017;Bharath et al 2019). Samples were acquired from different parts of the oral region, including from whole teeth (Srivastava et al 2017;Lim et al 2019), dentin or dental pulp (Zapico and Ubelaker 2013;Kholief et al 2017;Dutta et al 2017;Chowdhury et al 2018), epithelial cells from toothbrush, intraoral, dental prosthesis (Reddy et al 2011;Nakanishi et al 2015;Bharath et al 2019), and saliva from dental prosthesis (Chauhan et al 2016). Three studies mention the age of sampled subjects who were 19-81 years old (Zapico and Ubelaker 2013;Kholief et al 2017;Bharath et al 2019).…”

“…Samples were acquired from different parts of the oral region, including from whole teeth (Srivastava et al 2017;Lim et al 2019), dentin or dental pulp (Zapico and Ubelaker 2013;Kholief et al 2017;Dutta et al 2017;Chowdhury et al 2018), epithelial cells from toothbrush, intraoral, dental prosthesis (Reddy et al 2011;Nakanishi et al 2015;Bharath et al 2019), and saliva from dental prosthesis (Chauhan et al 2016). Three studies mention the age of sampled subjects who were 19-81 years old (Zapico and Ubelaker 2013;Kholief et al 2017;Bharath et al 2019). The other studies do not mention the subject's age (Reddy et al 2011;Nakanishi et al 2015;Chauhan et al 2016;Srivastava et al 2017;Dutta et al 2017;Chowdhury et al 2018;Lim et al 2019).…”

“…DNA extraction was performed by Chelex-100 (Reddy et al 2011;Bharath et al 2019), silica-based (Zapico and Ubelaker 2013;Chauhan et al 2016;Kholief et al 2017), silica-coated magnetic particle (Nakanishi et al 2015), or phenol-chloroform method (Srivastava et al 2017;Dutta et al 2017;Chowdhury et al 2018;Lim et al 2019). The methods of analysis in the studies included the amelogenin method (Srivastava et al 2017;Dutta et al 2017;Chowdhury et al 2018;Lim et al 2019), amelogenin with β-actin (Zapico and Ubelaker 2013), amelogenin without copy number variation (CNV) region (Nakanishi et al 2015), amelogenin and 15 CODIS loci (Chauhan et al 2016), SRY (Reddy et al 2011;Bharath et al 2019), and combined DYS14 along with SRY (Kholief et al 2017). DNA amplification was conducted using PCR (Lim et al 2019).…”