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
Citations
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Rapid techniques for DNA extraction from routinely processed archival tissue for use in PCR.
R Sepp,I Szabó,H Uda,H Sakamoto +3 more
TL;DR: Chelex boiling, proteinase K digestion followed by Chelex boiling produce suitable templates for the PCR from a large variety of paraffin wax embedded tissues, and could be useful, especially in the routine processing of large amounts of material.
Journal ArticleDOI
Degenerate and Nested PCR: a Highly Sensitive and Specific Method for Detection of Human Papillomavirus Infection in Cutaneous Warts
Catherine A. Harwood,Patricia J. Spink,T. Surentheran,Irene M. Leigh,Ethel Michele De Villiers,Jane M. McGregor,Charlotte M. Proby,Judith Breuer +7 more
TL;DR: A highly sensitive and comprehensive degenerate PCR methodology for detection and genotyping of HPV from the skin is established and a diverse spectrum of multiple HPV types in cutaneous warts from transplant recipients is demonstrated.
Journal ArticleDOI
PCR in diagnosis of infection: detection of bacteria in cerebrospinal fluids.
TL;DR: Current studies concerning detection of Chlamydia pneumoniae in CSF obtained from patients with multiple sclerosis by using PCR provide a good example for discussion of use of the PCR assay in diagnosis.
Journal ArticleDOI
Rapid reverse transcription-PCR detection of hepatitis C virus RNA in serum by using the TaqMan fluorogenic detection system.
T Morris,B Robertson,M Gallagher +2 more
TL;DR: TaqMan compared favorably with nested PCR with key advantages of speed, increased throughput, and decreased opportunity for false-positive results because of elimination of second-round amplification.
Journal ArticleDOI
Occult Hepatitis B Virus Infection and Clinical Outcomes of Patients with Chronic Hepatitis C
TL;DR: The data suggest that occultHBV infection does not have clinical significance in chronic hepatitis C patients residing in areas where HBV infection is endemic.
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.