Fully integrated, fully automated generation of short tandem repeat profiles

Investigative Genetics

Richard F Selden¹  James W Schumm¹  Luke Palombo¹  Sameer Vasantgadkar¹  Catherine Hogan¹  Rosemary S Turingan¹  Eugene Tan¹ 

[1] NetBio, 830 Winter Street, Waltham, MA 02451, USA

关键词: Short tandem repeat;    Rapid DNA analysis;    Polymerase chain reaction;    Microfluidics;    CODIS;    Automation;   

Others  :  801469
DOI  :  10.1186/2041-2223-4-16

 received in 2013-05-25, accepted in 2013-07-18,  发布年份 2013

【 摘 要 】

Background

The generation of short tandem repeat profiles, also referred to as ‘DNA typing,’ is not currently performed outside the laboratory because the process requires highly skilled technical operators and a controlled laboratory environment and infrastructure with several specialized instruments. The goal of this work was to develop a fully integrated system for the automated generation of short tandem repeat profiles from buccal swab samples, to improve forensic laboratory process flow as well as to enable short tandem repeat profile generation to be performed in police stations and in field-forward military, intelligence, and homeland security settings.

Results

An integrated system was developed consisting of an injection-molded microfluidic BioChipSet cassette, a ruggedized instrument, and expert system software. For each of five buccal swabs, the system purifies DNA using guanidinium-based lysis and silica binding, amplifies 15 short tandem repeat loci and the amelogenin locus, electrophoretically separates the resulting amplicons, and generates a profile. No operator processing of the samples is required, and the time from swab insertion to profile generation is 84 minutes. All required reagents are contained within the BioChipSet cassette; these consist of a lyophilized polymerase chain reaction mix and liquids for purification and electrophoretic separation.

Profiles obtained from fully automated runs demonstrate that the integrated system generates concordant short tandem repeat profiles. The system exhibits single-base resolution from 100 to greater than 500 bases, with inter-run precision with a standard deviation of ±0.05 - 0.10 bases for most alleles. The reagents are stable for at least 6 months at 22°C, and the instrument has been designed and tested to Military Standard 810F for shock and vibration ruggedization. A nontechnical user can operate the system within or outside the laboratory.

Conclusions

The integrated system represents the first generation of a turnkey approach to short tandem repeat profiling and has the potential for use in both the field (for example, police booking stations, the battlefield, borders and ports) and the forensic laboratory.

【 授权许可】

   
2013 Tan et al.; licensee BioMed Central Ltd.

【 图 表 】

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