Bioassay for specific DNA sequences using a non-radioactive probe |
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| Authors: | J L Hartley M Berninger J A Jessee F R Bloom G F Temple |
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| Affiliation: | 1. Health Sciences Research Centre, Faculty of Health, Student Research Committee, Mazandaran University of Medical Sciences, Sari, Iran;2. Department of Environmental Engineering, Faculty of Civil Engineering, Yildiz Technical University, Davutpasa Campus, 34220, Esenler, Istanbul, Turkey;3. Faculty of Health and Health Sciences Research Center, Department of Environmental Health Engineering, Mazandaran University of Medical Sciences, Sari, Iran;4. CEIT, Unit of Environmental Engineering, Paseo Manuel de Lardizábal 15, 20018 San Sebastian, Spain;5. IKERBASQUE, Basque Foundation for Science, María Díaz de Haro 3, 48013 Bilbao, Spain;6. Department of Chemistry, Indian Institute of Technology Roorkee, 247667, India;7. Department of Applied Chemistry, University of Johannesburg, Johannesburg, South Africa;1. State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu Province 730046, PR China;2. College of Animal Science and Technology, Anhui Agricultural University, Hefei, Anhui Province 230036, PR China;3. College of Veterinary Medicine, Hunan Agricultural University, Changsha, Hunan Province 410128, PR China;4. Department of Medical Zoology, Mie University School of Medicine, Mie 514-8507, Japan;5. Department of Veterinary Parasitology, Jeju National University College of Veterinary Medicine, Jeju 690-756, Republic of Korea;6. Department Parasitology and Tropical Medicine, Institute of Health Sciences, Gyeongsang National University School of Medicine, Jinju 52727, Republic of Korea;7. Department of Parasitology, National Institute of Infectious Diseases, Tokyo 162-8640, Japan;1. Division of Developmental Cognitive Neuroscience, Institute of Development, Aging and Cancer, Tohoku University, Sendai, Japan;2. Division of Medical Neuroimaging Analysis, Department of Community Medical Supports, Tohoku Medical Megabank Organization, Tohoku University, Sendai, Japan;3. Department of Radiology and Nuclear Medicine, Institute of Development, Aging and Cancer, Tohoku University, Sendai, Japan;4. Department of Functional Brain Imaging, Institute of Development, Aging and Cancer, Tohoku University, Sendai, Japan;5. Human and Social Response Research Division, International Research Institute of Disaster Science, Tohoku University, Sendai, Japan;6. Smart Ageing International Research Center, Institute of Development, Aging and Cancer, Tohoku University, Sendai, Japan;7. Japan Society for the Promotion of Science, Tokyo, Japan;8. Faculty of Medicine, Tohoku University, Sendai, Japan;1. Manchester Institute of Biotechnology (MIB), School of Chemistry, The University of Manchester, Manchester, United Kingdom;2. Discovery Biology, Discovery Sciences, IMED Biotech Unit, AstraZeneca, Cambridge, United Kingdom |
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| Abstract: | A novel method for detecting specific DNA sequences is described. The method uses a non-radioactive DNA probe, called a probe-vector, that can transform competent Escherichia coli cells at high efficiency only when it has hybridized to a specific DNA target, thus forming a circular, double-stranded, plasmid-like molecule. The probe-vector carries a plasmid origin of replication and a gene that confers antibiotic resistance on transformed E. coli. The output of the assay--colored bacterial colonies on an agar plate--is quantitative and proportional over a wide range of target concentrations. The utility of the probe-vector method for detecting hepatitis B virus (HBV) DNA in human serum is demonstrated. The assay can detect as little as 0.1 pg HBV DNA. The presence of an internal standard monitors DNA recovery and E. coli transformation efficiency for each sample. The assay has the potential to simultaneously measure the DNA of two or more pathogens within the same clinical sample. |
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