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A novel polymerase chain reaction method for detection of human immunodeficiency virus in dried blood spots on filter paper.
- Source :
-
Journal of clinical microbiology [J Clin Microbiol] 1992 Nov; Vol. 30 (11), pp. 2887-92. - Publication Year :
- 1992
-
Abstract
- A method for detection of proviral human immunodeficiency virus DNA in dried blood spots on filter paper by direct polymerase chain reaction (PCR) has been developed. To develop the method, a standard system was used which was prepared from cells each containing a single integrated provirus and titrated with normal donor blood. This rapid procedure provides virtually quantitative yields of nuclear DNA and exploits most of the standard methodology described for blood specimens. A nested PCR using SK38-SK39 gag as the internal primer pair was also designed; this PCR detected a single copy of provirus per filter at near theoretical frequency with SK19 probe. The utility of the procedure was demonstrated with clinical specimens. Blood spot filters from human immunodeficiency virus-infected and uninfected individuals were readily and unequivocally discriminated. The method is designed for ultimate use with large (1.5-ml) sample preparation tubes that are compatible as PCR tubes with thermal cyclers. This will permit convenient, direct single-tube PCR of dried blood specimens on filters. It should be adaptable to analysis of dried blood spots for a variety of infectious or genetic diseases.
- Subjects :
- Base Sequence
Blood Specimen Collection
Child
Child, Preschool
DNA, Viral blood
DNA, Viral genetics
Evaluation Studies as Topic
Genes, gag
HIV Infections blood
HIV Infections microbiology
Humans
Infant
Infant, Newborn
Molecular Sequence Data
Paper
Polymerase Chain Reaction standards
Reference Standards
HIV genetics
HIV isolation & purification
HIV Infections diagnosis
Polymerase Chain Reaction methods
Subjects
Details
- Language :
- English
- ISSN :
- 0095-1137
- Volume :
- 30
- Issue :
- 11
- Database :
- MEDLINE
- Journal :
- Journal of clinical microbiology
- Publication Type :
- Academic Journal
- Accession number :
- 1452659
- Full Text :
- https://doi.org/10.1128/jcm.30.11.2887-2892.1992