A highly sensitive chemiluminescent reverse transcriptase assay for human immunodeficiency virus
| Autor: | Youko Nagai-Fujii, Hideo Misaki, Kenji Arai, Kouichi Sano, Fumitomo Odawara, Hiroshi Azuma, Takehiro Kohno, Hideki Abe, Sharad Mohan, Kenji Ikebuchi, Shigeyuki Imamura, Hisami Ikeda |
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| Rok vydání: | 2002 |
| Předmět: |
T-Lymphocytes
HIV Infections Sensitivity and Specificity Colorimetry (chemical method) Virus law.invention Retrovirus law Virology Deoxyribonuclease I Humans Cells Cultured Chemiluminescence chemistry.chemical_classification biology Reverse Transcriptase Polymerase Chain Reaction biology.organism_classification Nucleotidyltransferase Molecular biology HIV Reverse Transcriptase Reverse transcriptase Enzyme chemistry Biochemistry Cell culture Luminescent Measurements HIV-1 RNA Viral Colorimetry |
| Zdroj: | Journal of Virological Methods. 106:115-124 |
| ISSN: | 0166-0934 |
| DOI: | 10.1016/s0166-0934(02)00142-8 |
| Popis: | A simple and highly sensitive reverse transcriptase (RT) assay was developed by combining a previously reported non-radioisotopic RT assay with the use of a template-primer-immobilized microplate, an enzyme capture protocol, product digestion and a chemiluminescent substrate. The assay was able to detect directly the RT activity in serum samples, plasma and cell culture medium without the need for concentration and extraction of the enzyme. The assay was able to detect RT activity equivalent to 100 virions/ml of HIV-1. These results suggest that this highly sensitive chemiluminescent RT assay can be used not only for virological investigation but also for routine screening of biopharmaceuticals. |
| Databáze: | OpenAIRE |
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