Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz
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SHR Neuro Krebs Kardio Lipid Stoffw Microb
Freystetter, A; Paar, C; Stekel, H; Berg, J.
Armored DNA in recombinant Baculoviruses as controls in molecular genetic assays.
Appl Microbiol Biotechnol. 2017; 101(19):7259-7269
Doi: 10.1007/s00253-017-8436-9
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- Führende Autor*innen der Med Uni Graz
- Berg Jörg
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- Abstract:
- The widespread use of molecular PCR-based assays in analytical and clinical laboratories brings about the need for test-specific, stable, and reliable external controls (EC) as well as standards and internal amplification controls (IC), in order to arrive at consistent test results. In addition, there is also a growing need to produce and provide stable, well-characterized molecular controls for quality assurance programs. In this study, we describe a novel approach to generate armored double-stranded DNA controls, which are encapsulated in baculovirus (BV) particles of the species Autographa californica multiple nucleopolyhedrovirus. We used the well-known BacPAK™ Baculovirus Expression System (Takara-Clontech), removed the polyhedrin promoter used for protein expression, and generated recombinant BV-armored DNAs. The obtained BV-armored DNAs were readily extracted by standard clinical DNA extraction methods, showed favorable linearity and performance in our clinical PCR assays, were resistant to DNase I digestion, and exhibited marked stability in human plasma and serum. BV-armored DNA ought to be used as ECs, quantification standards, and ICs in molecular assays, with the latter application allowing for the entire monitoring of clinical molecular assays for sample adequacy. BV-armored DNA may also be used to produce double-stranded DNA reference materials for, e.g., quality assurance programs. The ease to produce BV-armored DNA should make this approach feasible for a broad spectrum of molecular applications. Finally, as BV-armored DNAs are non-infectious to mammals, they may be even more conveniently shipped than clinical specimen.
- Find related publications in this database (using NLM MeSH Indexing)
- Amino Acid Sequence -
- Baculoviridae - genetics
- Cloning, Molecular -
- DNA, Viral - genetics
- Deoxyribonuclease I - metabolism
- Gene Expression Regulation, Viral -
- Plasmids - genetics
- Sequence Analysis, DNA -
- Find related publications in this database (Keywords)
- Baculovirus
- PCR
- Armored double-stranded DNA
- DNA control
- Internal control
- External control
- Autographa californica