DNA detection in tooth exposed to different temperatures: An in vitro study

Devaraju, Rama Raju; Gantala, Ramlal; Ambati, Madhavi; Vemula, Arjun; Kubbi, Jitender Reddy; Gotoor, Srikanth

doi:10.4103/0972-1363.155681

Cited by 5 publications

(4 citation statements)

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“…This may suggest that the phenol-chloroform method is still reliable and can produce sufficient DNA from the burnt and degraded tooth samples. As stated by previous studies, the DNA yielded from tooth can be affected by condition of tooth and chronological age of individual 12,13 . Chronological age has positive effect on DNA yield because mature teeth are more mineralised and less porous, contributing to higher resistance to decomposition and more protection for endogenous DNA 13 .…”

Section: Genomic Dna Extractionmentioning

confidence: 77%

Application of Two Sex Markers by Nested PCR for Gender Determination

Jonathan Jun-Yong Lim,

Mohd Fadhli Khamis,

Nur Haslindawaty Binti Abd Rashid

2021

Indian Journal of Forensic Medicine & Toxicology

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Sex determination is one of the basic components in victim identification. There are many available methods, namely forensic anthropology method and conventional DNA typing method. In this study, nested PCR technique was employed in sex typing of burnt teeth through amelogenin (AMEL) and sex-determining region Y (SRY) markers. In this study, 17 teeth samples were burnt at temperatures that ranged from 100°C to 500°C for 2 min -10 min. The whole tooth was used for DNA extraction by phenol-chloroform method. Accurate sex determination was achieved in 13 samples by both AMEL and SRY markers. The SRY marker achieved higher sensitivity as compared to AMEL marker. The sensitivity of both markers was improved consequent to nested PCR. Factors such as degraded DNA materials and the presence of tooth caries greatly affect sex typing results. Results showed that nested PCR proved to be a good method to amplify highly degraded DNA material as it greatly increased the DNA copy, and thus increased the possibility of sex typing.

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Section: Genomic Dna Extractionmentioning

confidence: 77%

Application of Two Sex Markers by Nested PCR for Gender Determination

Jonathan Jun-Yong Lim,

Mohd Fadhli Khamis,

Nur Haslindawaty Binti Abd Rashid

2021

Indian Journal of Forensic Medicine & Toxicology

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“…*Abbreviations = M -Male; F -Female the external environment in caries tooth. Devaraju et al (2014) has revealed tooth with more caries has less amount of DNA. Other previous studies stated that dental disease includes dental caries and periodontitis have a negative impact on the human DNA content of teeth (Higgins et al 2011;Dogan et al 2017).…”

Section: Discussionmentioning

confidence: 99%

“…The DNA in tooth is also preserved in the sealed pulp cavity and protected from harsh environmental conditions , making it a reliable source for sex determination even after decomposition. A study found that DNA material of 39.5 µg/mL to 60 µg/mL can be obtained from the tooth samples exposed to temperatures of 100°C to 300°C for 10 to 15 minutes, proving tooth to be suitable as forensic evidence material (Devaraju et al 2014;Praveen Kumar & Aswath 2016). Since sex typing was determined on untreated tooth samples using amelogenin gene (Praveen Kumar & Aswath 2016), and DNA has been successfully extracted from incinerated tooth followed by sex typing using Y-specific actin gene (Shrishail et al 2011), thus, this study aims to conduct sex typing on incinerated tooth at different temperatures and duration using conventional sex marker, amelogenin gene through nested PCR method for sex determination.…”

Section: Introductionmentioning

confidence: 99%

Identification of Amelogenin Gene on Burnt Teeth Samples through Nested Polymerase Chain Reaction Amplification for Sex Identification

Lim

Khamis

Rashid

2019

JSKM

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Sex determination is one of the basic components in victim identification. This study aims to ascertain the sex of an individual from burnt teeth samples exposed at different temperature and time through nested polymerase chain reaction (PCR) on the amelogenin (AMEL) sex marker, to calculate the specificity and sensitivity, and to compare with previous relevant studies. A total of 17 teeth samples was subjected to burning at different temperatures ranging from 100°C to 500°C, at 2 to 10 minutes. The whole tooth was used for deoxyribonucleic acid (DNA) extraction by phenol-chloroform method. All samples were quantified for DNA concentration and then analyzed with nested PCR using two pairs of AMEL primer and results of sex typing were recorded. Out of 17 samples, genomic DNA extracted from 6 samples have concentrations ranging from 27.3-130.6 ng/µL. Nested PCR could amplify 16 samples for AMEL gene. Sex typing using AMEL gene showed 76.47% accuracy. Sensitivity of AMEL primer was increased from 6.67% to 63.64% using nested PCR technique; specificity of both external and internal primer was reported at 100%. Nested PCR of AMEL gene proved to be a suitable method for unequivocal determination of sex from degraded DNA samples.

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“…It has generally been reported that dental DNA can withstand temperatures up to 400 °C for one hour but that its quantity and quality are superior after exposure to lower temperatures for shorter time periods (1–20 min), facilitating forensic identification 9 . Only a few studies have studied teeth exposed for more than 15 min at temperatures above 90 °C 16 , 17 , 18 , 19 . Most researchers have reported that it is very difficult to extract DNA from teeth after their exposure to temperatures of 200–400 °C 18 , 19 , 20 , and no consensus has been reached on the degree of cremation at which teeth will still yield nuclear DNA signals 7 .…”

Section: Introductionmentioning

confidence: 99%

DNA degradation in human teeth exposed to thermal stress

Lozano-Peral

Rubio

Santos

et al. 2021

Sci Rep

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Human identification from burned remains poses a challenge to forensic laboratories, and DNA profiling is widely used for this purpose. Our aim was to evaluate the effect of temperature on DNA degradation in human teeth. Thirty teeth were exposed to temperatures of 100, 200, or 400 °C for 60 min. DNA was quantified by Real-Time qPCR (Quantifiler Human DNA Quantification Kit) and fluorescence spectroscopy (Qubit 3.0 Fluorometer). DNA degradation was evaluated by using STR markers (AmpFLSTR Identifiler Plus PCR Amplification Kit) to determine the allele and locus dropout, inter-locus balance, and degradation slope (observed (Oa) to expected (Ea) locus peak height ratio against the molecular weight). Most of the genomic DNA was degraded between 100 °C and 200 °C. At 100 °C, locus dropout ratios showed significant differences between the largest loci (FGA, D7S820, D18S51, D16S539, D2S1338 and CSF1PO) and amelogenin. Inter-locus balance values significantly differed between all dye channels except between NED and PET. The dropout ratio between D18S51 (NED) and amelogenin (PET) can be recommended for the evaluation of DNA degradation. The Oa/Ea regression model can predict locus peak heights in DNA degradation (R2 = 0.7881). These findings may be useful to assess the reliability of DNA typing for human identification in teeth subjected to prolonged incineration.

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DNA detection in tooth exposed to different temperatures: An in vitro study

Cited by 5 publications

References 0 publications

Application of Two Sex Markers by Nested PCR for Gender Determination

Application of Two Sex Markers by Nested PCR for Gender Determination

Identification of Amelogenin Gene on Burnt Teeth Samples through Nested Polymerase Chain Reaction Amplification for Sex Identification

DNA degradation in human teeth exposed to thermal stress

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