Journal ArticleDOI
Avoiding false positives with PCR
S Kwok,Russell Higuchi +1 more
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TLDR
The exquisite sensitivity of the polymerase chain reaction means DNA contamination can ruin an entire experiment and adherence to a strict set of protocols can avoid disaster.Abstract:
The exquisite sensitivity of the polymerase chain reaction means DNA contamination can ruin an entire experiment. Tidiness and adherence to a strict set of protocols can avoid disaster.read more
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High-Resolution Hepatitis C Virus Subtyping Using NS5B Deep Sequencing and Phylogeny, an Alternative to Current Methods
Josep Quer,Josep Quer,Josep Gregori,Josep Gregori,Francisco Rodriguez-Frias,Francisco Rodriguez-Frias,Maria Buti,Maria Buti,Antonio Madejón,Sofía Pérez-del-Pulgar,Damir Garcia-Cehic,Rosario Casillas,Maria Blasi,Maria Homs,David Tabernero,Miguel Álvarez-Tejado,Jose Manuel Muñoz,Maria Cubero,Andrea Caballero,Jose Antonio delCampo,Esteban Domingo,Esteban Domingo,Irene Belmonte,L. Nieto,Sabela Lens,Paloma Muñoz-de-Rueda,Paloma Sanz-Cameno,Silvia Sauleda,Marta Bes,Jordi Gómez,Jordi Gómez,Carlos Briones,Carlos Briones,Celia Perales,Celia Perales,Julie Sheldon,Julie Sheldon,Lluis Castells,Lluis Castells,L Viladomiu,Javier Salmerón,Angela Ruiz-Extremera,Rosa Quiles-Pérez,Ricardo Moreno-Otero,Rosario López-Rodríguez,Helena Allende,Manuel Romero-Gómez,Jaume Guardia,Jaume Guardia,Rafael Esteban,Rafael Esteban,Javier García-Samaniego,Xavier Forns,Juan Ignacio Esteban,Juan Ignacio Esteban +54 more
TL;DR: It is concluded that deep sequencing is more efficient for HCV subtyping than currently available methods and allows qualitative identification of mixed infections and may be more helpful with respect to informing treatment strategies with new DAA-containing regimens across allHCV subtypes.
Journal ArticleDOI
Tumor cells of hairy cell leukemia express multiple clonally related immunoglobulin isotypes via RNA splicing.
Francesco Forconi,Surinder S. Sahota,Donatella Raspadori,C I Mockridge,Francesco Lauria,Freda K. Stevenson +5 more
TL;DR: Derivation of multiple isotypes from individual cells was demonstrated by analyzing transcripts in single sorted cells from one patient, with evidence for coexistence of isotype variants in 10 of 10 cells, indicating that clonally related multiple isotype coexist in single HCs.
Journal ArticleDOI
Disseminated tumour cells. Their detection and significance for prognosis of gastrointestinal and pancreatic carcinomas.
Ilka Vogel,Holger Kalthoff +1 more
TL;DR: The accuracy of the detection methods and the prognostic value of detecting disseminated tumour cells in the bone marrow, blood and peritoneal lavage of patients with colorectal, gastric and pancreatic carcinomas are discussed.
Journal ArticleDOI
Clonal rearrangements in childhood and adult precursor B acute lymphoblastic leukemia : a comparative polymerase chain reaction study using multiple sets of primers.
Aihong Li,Richard Rosenquist,Erik Forestier,Dan Holmberg,Jack Lindh,Eva Löfvenberg,Göran Roos +6 more
TL;DR: A common heterogeneity was demonstrated in different compartments reflecting ongoing clonal evolution, which can make detection of minimal residual disease (MRD) in ALL troublesome and it is suggested that a minimum of three targets should be used to minimise false‐negative results.
Journal ArticleDOI
Human herpesvirus‐6: A survey of presence and distribution of genomic sequences in normal brain and neuroglial tumors
Mario Luppi,P. Barozzi,Antonio Maiorana,Roberto Marasca,Raffaella Trovato,R. A. Fano,Luca Ceccherini-Nelli,Giuseppe Torelli +7 more
TL;DR: Molecular evidence of a wide distribution of HHV‐6 infection in the brain tissues of a high proportion of subjects, both in normal and in impaired immunity is provided, arguing against a major role of this herpesvirus in the pathogenesis of primary brain tumors of neuroglial origin in immunocompetent subjects.
References
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Journal ArticleDOI
Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase
Randall Keichi Saiki,David H. Gelfand,Susanne Stoffel,Stephen J. Scharf,Russell Higuchi,Glenn Thomas Horn,Kary B. Mullis,Henry A. Erlich +7 more
TL;DR: A thermostable DNA polymerase was used in an in vitro DNA amplification procedure, the polymerase chain reaction, which significantly improves the specificity, yield, sensitivity, and length of products that can be amplified.
Book ChapterDOI
Specific synthesis of DNA in vitro via a polymerase-catalyzed chain reaction.
Kary B. Mullis,Fred A. Faloona +1 more
TL;DR: A method whereby a nucleic acid sequence can be exponentially amplified in vitro is described in the chapter, and the possibility of utilizing a heat-stable DNA polymerase is explored so as to avoid the need for addition of new enzyme after each cycle of thermal denaturation.
Journal ArticleDOI
DNA amplification for direct detection of HIV-1 in DNA of peripheral blood mononuclear cells
Chin-Yih Ou,Shirley Kwok,Sheila W. Mitchell,David Henry Mack,John J. Sninsky,John W. Krebs,Paul M. Feorino,Donna T. Warfield,Gerald Schochetman +8 more
TL;DR: This method of DNA amplification made it possible to obtain results within 3 days, whereas virus isolation takes up to 3 to 4 weeks, and may therefore be used to complement or replace virus isolation as a routine means of determining HIV-1 infection.
Journal ArticleDOI
DNA typing from single hairs
TL;DR: Three different means of DNA typing are used for the determination of amplified DNA fragment length differences, hybridization with allele-specific oligonucleotide probes, and direct DNA sequencing on single human hairs to detect genetically variable mitochondrial and nuclear DNA sequences.
Journal ArticleDOI
Amplification and analysis of DNA sequences in single human sperm and diploid cells
Honghua Li,Ulf B. Gyllensten,Xiangfeng Cui,Randall Keichi Saiki,Henry A. Erlich,Norman Arnheim +5 more
TL;DR: The use of the polymerase chain reaction for analysing DNA sequences in individual diploid cells and human sperm shows that two genetic loci can be co-amplified from a single sperm, which may allow the analysis of previously inaccessible genetic phenomena.