Document Details

Document Type : Article In Journal 
Document Title :
Development and validation of a novel multiplexed DNA analysis system, InnoTyper® 21
Development and validation of a novel multiplexed DNA analysis system, InnoTyper® 21
 
Document Language : English 
Abstract : We report here a novel multiplexed DNA analysis system consisting of 20 Alu markers and Amelogenin for analysis of highly degraded forensic biological samples. The key to the success of the system in obtaining results from degraded samples is the primer design yielding small amplicon size (60–125 bp) for all 20 markers. The markers included in the InnoTyper1 21 system are bi-allelic, having two possible allelic states (insertion or null) and thus termed INNULs. The markers are short interspersed nuclear elements (SINEs), a category of retrotransposable elements (REs) which are non-coding genomic DNA repeat sequences, or “mobile insertion elements,” comprising approximately 40% of the human genome. Alu elements are primate specific SINEs that have reached a copy number in excess of one million in the human genome, which makes these markers highly sensitive and desirable for forensic samples with extremely degraded DNA. Until now however, due to the inherent size difference associated with insertion and no insertion alleles, the use of Alu REs has not been practical for forensic applications. The novel primer design described herein has allowed the development of a multiplexed Alu system yielding fragment sizes amenable to degraded DNA samples, as frequently encountered in missing persons cases or forensic samples such as hair shafts. Although use of Alus in human identity has been studied using single marker amplification and reported before, we report for the first time development and validation of a system with multiplexed RE markers. Studies performed include PCR optimization, species specificity, sensitivity, degradation and inhibition, precision and accuracy, nonprobative samples, mixture, and population database studies. A population study using 592 samples including five populations was performed using InnoTyper 21. The data indicated the random match probability for the combination of these 20 Alu markers was greater than 1 in 3.8 million for the populations studied, indicating the greater statistical power of these autosomal nuclear DNA markers over haplotype systems typically used in such degraded samples. Results demonstrate the system is successful in obtaining results from highly degraded DNA. A sensitivity study performed demonstrated at least 95% recovery of alleles from as low as 50 pg of total input DNA, and partial profiles from as low as 25 pg. This study has demonstrated that the bi-allelic INNULs in the InnoTyper 21 system provide a sensitivity of detection and a power of discrimination that makes them useful for human identification of extremely degraded samples. 
ISSN : 1872-4973 
Journal Name : Forensic Science International: Genetics 
Volume : 29 
Issue Number : 1 
Publishing Year : 1438 AH
2017 AD 
Article Type : Article 
Added Date : Thursday, June 1, 2017 

Researchers

Researcher Name (Arabic)Researcher Name (English)Researcher TypeDr GradeEmail
Brown HiromiHiromi, Brown ResearcherDoctorate 
Thompson RobynRobyn, Thompson ResearcherDoctorate 
Murphy GinaGina, Murphy ResearcherDoctorate 
Peters DixieDixie, Peters ResearcherDoctorate 
La Rue BobbyBobby, La Rue ResearcherDoctorate 
King JonathanJonathan, King ResearcherDoctorate 
Montgomery Anne HAnne H, Montgomery ResearcherDoctorate 
Carroll MarionMarion, Carroll ResearcherDoctorate 
Baus JamesJames, Baus ResearcherDoctorate 
Sinha SidSid, Sinha ResearcherDoctorate 
Wendt R. FrankFrank, Wendt R.ResearcherDoctorate 
Song BingBing, Song ResearcherDoctorate 
Chakraborty RanajitRanajit, Chakraborty ResearcherDoctorate 
Budowle BruceBruce, Budowle ResearcherDoctorate 
Sinha SudhirSudhir, Sinha InvestigatorDoctorate 

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