Efficient amplification with NASBA® of hepatitis B virus, herpes simplex virus and methicillin resistant Staphylococcus aureus DNA
| Autor: | Dianne van Strijp, Fokke Venema, Carine Zintilini, Corinne Jay, Paul van de Wiel, Birgit Deiman, Saskia Vermeer |
|---|---|
| Rok vydání: | 2008 |
| Předmět: |
Hepatitis B virus
Staphylococcus aureus Nucleic acid amplification technique Biology biology.organism_classification medicine.disease_cause Sensitivity and Specificity Virology NASBA Virus Herpes simplex virus Orthohepadnavirus Hepadnaviridae DNA Viral medicine Simplexvirus Methicillin Resistance Primer (molecular biology) Nucleic Acid Amplification Techniques Self-Sustained Sequence Replication DNA Primers |
| Zdroj: | Journal of Virological Methods. 151:283-293 |
| ISSN: | 0166-0934 |
| Popis: | A new mechanism is described for DNA amplification using nucleic acid sequence-based amplification (NASBA) including a restriction enzyme digestion and P1 primer binding directly upstream of the digestion. For hepatitis B virus (HBV), herpes simplex virus (HSV) and methicillin resistant Staphylococcus aureus (MRSA) DNA, which all show very poor amplification with normal NASBA, assay sensitivity was improved by a factor 100-1000 when restriction enzyme digestion was performed prior to amplification. For the quantitative HBV assay, in combination with the NucliSENS Extractor (bioMerieux, Boxtel, The Netherlands), a 95% target detection rate of 242 WHO IU/ml and 50% detection rate of 35 WHO IU/ml was achieved. The lowest detectable HBV concentration was 10 WHO IU/ml. HBV DNA could be quantified with an algorithm comparable to that used for RNA quantitation and by using a two step approach a dynamic range of 10(2)-10(9)WHO IU/ml (>6 log) was shown to be quantifiable. For the qualitative HSV assay, in combination with the NucliSENS miniMAG (bioMerieux, Boxtel, The Netherlands), the 95% detection rate was determined to be 84 and 138 copies/isolation for HSV 1 and HSV 2, respectively, which corresponds to approximately 10 copies per amplification for both targets. For MRSA, the limit of detection was |
| Databáze: | OpenAIRE |
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