Quantification of HIV-1 Using Multiple Competitors in a Single-Tube Assay
| Autor: | T. Vener, M. Axelsson, J. Albert, M. Uhlén, J. Lundeberg |
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| Jazyk: | angličtina |
| Rok vydání: | 1996 |
| Předmět: | |
| Zdroj: | BioTechniques, Vol 21, Iss 2, Pp 248-255 (1996) |
| Druh dokumentu: | article |
| ISSN: | 1940-9818 0736-6205 |
| DOI: | 10.2144/96212st01 |
| Popis: | Methods for quantification of human immunodeficiency virus type 1 (HIV-1) based on competitive PCR and fragment analysis have been developed. Samples containing HIV-1 DNA and known amounts of three cloned competitors were co-amplified by PCR with semi-nested primers. The competitor DNAs contained the same long terminal repeat primer binding sequences as the wild-type DNA, but they are different in internal sequences and length. One of the inner primers was fluorescent-labeled to allow discrimination between the wild-type DNA and the three competitors by fragment analysis using a standard automated sequencer. A calibration curve using the peak area of the three competitors enabled accurate determination of target amount with minimal variations. The method presented here can be used for quantification of HIV-1 in clinical samples and will be useful for monitoring disease progression and treatment effects. |
| Databáze: | Directory of Open Access Journals |
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