Biomaterial Database

Typing human T-cell lymphotropic virus (HTLV-I and HTLV-II) by nested polymerase chain reaction: application to clinical specimens. 1995

A Vallejo, and A García-Sáiz

Servicio de Diagnóstico y Referencia, Instituto de Salud Carlos III, Madrid, Spain.

Human T-cell lymphotropic virus type I and II provirus DNA was detected by polymerase chain reaction (PCR). MT-2 (HTLV-I infected), C3/44 Mo (HTLV-II infected) cell lines and peripheral blood mononuclear cells (PBMNC) from HTLV seropositive samples were used. The procedure consists of first amplification which detects both HTLV-I and HTLV-II, and a second amplification (nested-PCR) to discriminate between the two viruses and to improve sensitivity. Optimal conditions of MgCl2 concentration and annealing temperature were found for maximal amplification and specificity. This method was used for the amplification of conserved regions of pol and env genes. 1.5 pg of MT-2 and 5 pg of C3/44 Mo cell line DNAs were detected using nested-PCR and liquid hybridization in the pol system. The env system could detect 1.5 pg of MT-2 and 1.5 pg of C3/44 Mo cell lines DNAs using nested-PCR and liquid hybridization. The pol system can type both HTLV-I and HTLV-II in only two steps without the use of type-specific radiolabeled probes. Furthermore, this method can detect and discriminate the two viruses in one step PCR using the primers used in the nested-PCR. Nevertheless, there is a decrease in sensitivity of 100-fold. The results of five seropositive samples confirmed by Western blot are compared with PCR. PCR typed one of these samples as HTLV-I and the rest as HTLV-II. This technique is useful in cases such as window period, perinatal studies and when serologic results are not satisfactory.

UI	MeSH Term	Description	Entries
D008969	Molecular Sequence Data	Descriptions of specific amino acid, carbohydrate, or nucleotide sequences which have appeared in the published literature and/or are deposited in and maintained by databanks such as GENBANK, European Molecular Biology Laboratory (EMBL), National Biomedical Research Foundation (NBRF), or other sequence repositories.	Sequence Data, Molecular,Molecular Sequencing Data,Data, Molecular Sequence,Data, Molecular Sequencing,Sequencing Data, Molecular
D011533	Proviruses	Duplex DNA sequences in eukaryotic chromosomes, corresponding to the genome of a virus, that are transmitted from one cell generation to the next without causing lysis of the host. Proviruses are often associated with neoplastic cell transformation and are key features of retrovirus biology.	Provirus
D002460	Cell Line	Established cell cultures that have the potential to propagate indefinitely.	Cell Lines,Line, Cell,Lines, Cell
D004279	DNA, Viral	Deoxyribonucleic acid that makes up the genetic material of viruses.	Viral DNA
D005069	Evaluation Studies as Topic	Works about studies that determine the effectiveness or value of processes, personnel, and equipment, or the material on conducting such studies.	Critique,Evaluation Indexes,Evaluation Methodology,Evaluation Report,Evaluation Research,Methodology, Evaluation,Pre-Post Tests,Qualitative Evaluation,Quantitative Evaluation,Theoretical Effectiveness,Use-Effectiveness,Critiques,Effectiveness, Theoretical,Evaluation Methodologies,Evaluation Reports,Evaluation, Qualitative,Evaluation, Quantitative,Evaluations, Qualitative,Evaluations, Quantitative,Indexes, Evaluation,Methodologies, Evaluation,Pre Post Tests,Pre-Post Test,Qualitative Evaluations,Quantitative Evaluations,Report, Evaluation,Reports, Evaluation,Research, Evaluation,Test, Pre-Post,Tests, Pre-Post,Use Effectiveness
D006801	Humans	Members of the species Homo sapiens.	Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D001483	Base Sequence	The sequence of PURINES and PYRIMIDINES in nucleic acids and polynucleotides. It is also called nucleotide sequence.	DNA Sequence,Nucleotide Sequence,RNA Sequence,DNA Sequences,Base Sequences,Nucleotide Sequences,RNA Sequences,Sequence, Base,Sequence, DNA,Sequence, Nucleotide,Sequence, RNA,Sequences, Base,Sequences, DNA,Sequences, Nucleotide,Sequences, RNA
D012680	Sensitivity and Specificity	Binary classification measures to assess test results. Sensitivity or recall rate is the proportion of true positives. Specificity is the probability of correctly determining the absence of a condition. (From Last, Dictionary of Epidemiology, 2d ed)	Specificity,Sensitivity,Specificity and Sensitivity
D014773	Virology	The study of the structure, growth, function, genetics, and reproduction of viruses, and VIRUS DISEASES.
D015153	Blotting, Western	Identification of proteins or peptides that have been electrophoretically separated by blot transferring from the electrophoresis gel to strips of nitrocellulose paper, followed by labeling with antibody probes.	Immunoblotting, Western,Western Blotting,Western Immunoblotting,Blot, Western,Immunoblot, Western,Western Blot,Western Immunoblot,Blots, Western,Blottings, Western,Immunoblots, Western,Immunoblottings, Western,Western Blots,Western Blottings,Western Immunoblots,Western Immunoblottings