A simple method for preparing synthetic controls for conventional and real-time PCR for the identification of endemic and exotic disease agents.
Identifieur interne : 001489 ( Ncbi/Checkpoint ); précédent : 001488; suivant : 001490A simple method for preparing synthetic controls for conventional and real-time PCR for the identification of endemic and exotic disease agents.
Auteurs : Greg Smith [Australie] ; Ina Smith ; Bruce Harrower ; David Warrilow ; Cheryl BletchlySource :
- Journal of virological methods [ 0166-0934 ] ; 2006.
Descripteurs français
- KwdFr :
- MESH :
- isolement et purification : Virus du SRAS.
- Humains, RT-PCR.
English descriptors
- KwdEn :
- MESH :
- isolation & purification : SARS Virus.
- methods : Reverse Transcriptase Polymerase Chain Reaction.
- Humans.
Abstract
Medical and veterinary diagnostic and public health laboratories world-wide are increasingly being called upon to introduce molecular diagnostic tests for both endemic and exotic diseases. This demand has accelerated following increasing terrorism fears. Ironically these same concerns have lead to tightening of both import and export controls preventing many laboratories, particularly those outside of the United States, from gaining access to positive control material. This in turn has prevented many laboratories from introducing much needed molecular diagnostic tests. We describe here a generic approach for preparing synthetic DNA or RNA control material for use in either TaqMan or conventional PCR assays. The production of synthetic controls using this approach does not require cloning or special equipment or facilities beyond that found in any laboratory performing molecular diagnostics. The approach significantly reduces the possibility of contamination or erroneously reporting false-positive reactions due to contamination from positive control material. Synthetic controls produced using this approach have been employed in all molecular diagnostic tests performed in our laboratory and are used irrespective of whether we possess the organism or not.
DOI: 10.1016/j.jviromet.2006.03.007
PubMed: 16677717
Affiliations:
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pubmed:16677717Le document en format XML
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<affiliation wicri:level="1"><nlm:affiliation>Public Health Virology, Centre for Public Health Sciences, Queensland Health Scientific Services, 39 Kessels Road, Coopers Plains, Brisbane, Qld 4108, Australia. greg_smith@health.qld.gov.au</nlm:affiliation>
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<author><name sortKey="Smith, Ina" sort="Smith, Ina" uniqKey="Smith I" first="Ina" last="Smith">Ina Smith</name>
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<keywords scheme="KwdFr" xml:lang="fr"><term>Humains</term>
<term>RT-PCR ()</term>
<term>Virus du SRAS (isolement et purification)</term>
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<keywords scheme="MESH" qualifier="isolement et purification" xml:lang="fr"><term>Virus du SRAS</term>
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<front><div type="abstract" xml:lang="en">Medical and veterinary diagnostic and public health laboratories world-wide are increasingly being called upon to introduce molecular diagnostic tests for both endemic and exotic diseases. This demand has accelerated following increasing terrorism fears. Ironically these same concerns have lead to tightening of both import and export controls preventing many laboratories, particularly those outside of the United States, from gaining access to positive control material. This in turn has prevented many laboratories from introducing much needed molecular diagnostic tests. We describe here a generic approach for preparing synthetic DNA or RNA control material for use in either TaqMan or conventional PCR assays. The production of synthetic controls using this approach does not require cloning or special equipment or facilities beyond that found in any laboratory performing molecular diagnostics. The approach significantly reduces the possibility of contamination or erroneously reporting false-positive reactions due to contamination from positive control material. Synthetic controls produced using this approach have been employed in all molecular diagnostic tests performed in our laboratory and are used irrespective of whether we possess the organism or not.</div>
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<name sortKey="Smith, Ina" sort="Smith, Ina" uniqKey="Smith I" first="Ina" last="Smith">Ina Smith</name>
<name sortKey="Warrilow, David" sort="Warrilow, David" uniqKey="Warrilow D" first="David" last="Warrilow">David Warrilow</name>
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<country name="Australie"><noRegion><name sortKey="Smith, Greg" sort="Smith, Greg" uniqKey="Smith G" first="Greg" last="Smith">Greg Smith</name>
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