| 1. |
- Alksnis, M., et al.
(författare)
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Use of synthetic oligodeoxyribonucleotides for type-specific identification of coxsackie B viruses
- 1989
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Ingår i: Molecular and Cellular Probes. - : Elsevier BV. - 0890-8508 .- 1096-1194. ; 3:2, s. 103-108
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Tidskriftsartikel (refereegranskat)abstract
- Synthetic oligodeoxyribonucleotides were used for type-specific identification of members of the coxsackie B virus group by nucleic acid hybridization. Two pairs of oligonucleotide chains were constructed based on nucleotide sequences in the VP1 regions of coxsackieviruses B3 and B4. Each labelled probe had a length of 24 nucleotides. The results showed that the oligonucleotide hybridized in a type-specific manner when assayed with extracts from cells infected with all different coxsackie B viruses. A method based on similar principles may thus be used for enterovirus typing.
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| 2. |
- Dahlén, P, et al.
(författare)
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Sensitive detection of genes by sandwich hybridization and time-resolved fluorometry
- 1987
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Ingår i: Molecular and Cellular Probes. - : Elsevier BV. - 0890-8508 .- 1096-1194. ; 1:2, s. 159-168
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Tidskriftsartikel (refereegranskat)abstract
- Europium has been used as a non-radioactive marker in immunoassays as this metal can be detected with high sensitivity by time-resolved fluorometry. In this work streptavidin labeled with europium was used to detect biotinylated probes in a sandwich nucleic-acid hybridization assay with microtitration strips as the solid phase. pBR 322 plasmids were detected with a sensitivity of 4 × 105 molecules. As the sample is added in solution in sandwich hybridization, fast and simple sample pre-treatment can be used without encountering background problems. The method was applied to test bacterial samples of uropathogenic Escherichia coli strains for the presence of the β-lactamase gene.
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| 3. |
- Jungell-Nortamo, A, et al.
(författare)
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Nucleic acid sandwich hybridization : enhanced reaction rate with magnetic microparticles as carriers
- 1988
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Ingår i: Molecular and Cellular Probes. - : Elsevier BV. - 0890-8508 .- 1096-1194. ; 2:4, s. 281-288
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Tidskriftsartikel (refereegranskat)abstract
- A method for the detection of nucleic acid hybrids using the sandwich hybridization technique with magnetic polystyrene microparticles as the solid support is described. The capture DNA is coupled to the polystyrene-hydroxy surface of the particles through p-toluenesulfonyl chloride activation. The use of microparticles results in a substantial increase in the reaction rate compared to filter hybridization, without decreasing the sensitivity of detection. Polyethylene glycol additionally enhances the reaction rate. The use of magnetic microparticles allows rapid and convenient collection of the formed hybrids.
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| 4. |
- Parkkinen, S, et al.
(författare)
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Sandwich hybridization in solution : a rapid method to screen HPV 16 DNA in cervical scrapes
- 1989
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Ingår i: Molecular and Cellular Probes. - : Elsevier BV. - 0890-8508 .- 1096-1194. ; 3:1, s. 1-11
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Tidskriftsartikel (refereegranskat)abstract
- A solution hybridization method is introduced as a rapid diagnostic method for demonstration of papillomavirus DNA in cervical scrapes. 32P-Labelled detector probe and the biotinylated capture probes were hybridized with DNA of the specimen after pretreatment by boiling in alkaline SDS. After 4 h of hybridization the hybrids were collected onto avidin coated beads and measured. The sensitivity of the method was 1–5 × 105 HPV 16 DNA molecules. Cervical carcinoma cell lines CaSki and SiHa were informative as to the sensitivity of the solution hybridization and the in situ hybridization methods. CaSki cells containing about 700 HPV 16 DNA copies per cell were positive by both methods. SiHa cells with one HPV 16 DNA copy per cell were positive by the sandwich assay but remained negative in the in situ test. A series of 126 cervical scrapes collected from consecutive patients participating in a follow-up study for cervical HPV infection were tested for HPV 16 DNA by both methods. The detection rate of the sandwich test was 19/126 (15%) and that of the in situ method 21/126 (17%) yielding 26 diagnoses altogether. Twelve of these were obtained by one method only. The results obtained by studying the cervical cell lines and repeated specimens taken from constantly HPV 16 positive patients suggested that the two methods can measure different types of infections and thus complement each other in the diagnosis of cervical HPV infections.
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