Abstract
The polymerase chain reaction was applied to the analysis of DNA contained in archival paraffin wax embedded material. DNA suitable for the reaction was obtained from these tissues by simple extraction methods, without previous dewaxing of tissue sections. When compared with unfixed material, the reaction efficiency was compromised, so that an increased number of amplification cycles were required to produce equivalent amounts of amplified product. This in turn led to an increase in amplification artefacts, which can be minimised by a simple modification of the standard reaction. Amplification of relatively large DNA fragments was not always successful, and it seems prudent to bear this in mind when designing oligonucleotide primers which are to be used for the amplification of archival material. The efficiency of the procedure can be improved by dividing the amplification cycles into two parts: this reduces the amount of reagent needed, is relatively simple and inexpensive, and can be performed in one working day.
MeSH Terms
Base Sequence
Culture Techniques
DNA, Viral/analysis
Electrophoresis, Agar Gel
Herpesvirus 4, Human/isolation & purification
Histological Techniques
Humans
Molecular Sequence Data
Nasopharyngeal Neoplasms/microbiology
Oligonucleotides
Paraffin
Polymerase Chain Reaction/methods
Chemicals
DNA, Viral
Oligonucleotides
Paraffin
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Coates P J
Department of Histopathology, St Bartholomew's Hospital, West Smithfield, London.
d'Ardenne A J
Khan G
Kangro H O
Slavin G
References (25)
25 references, click to expand
-
In vitro amplification of hepatitis B virus sequences from liver tumour DNA and from paraffin wax embedded tissues using the polymerase chain reaction.
J Clin Pathol. 1989 Aug;42(8):840-6
PMID: 2549102
-
DNA sequence and expression of the B95-8 Epstein-Barr virus genome.
Nature. 1984 Jul 19-25;310(5974):207-11
PMID: 6087149
-
Analysis of DNA extracted from formalin-fixed, paraffin-embedded tissues by enzymatic amplification and hybridization with sequence-specific oligonucleotides.
Biochem Biophys Res Commun. 1987 Feb 13;142(3):710-6
PMID: 3548717
-
Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia.
Science. 1985 Dec 20;230(4732):1350-4
PMID: 2999980
-
Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase.
Science. 1988 Jan 29;239(4839):487-91
PMID: 2448875
-
An improved method for prenatal diagnosis of genetic diseases by analysis of amplified DNA sequences. Application to hemophilia A.
N Engl J Med. 1987 Oct 15;317(16):985-90
PMID: 3657865
-
A potent inhibitor of Taq polymerase copurifies with human genomic DNA.
Nucleic Acids Res. 1988 Nov 11;16(21):10355
PMID: 3194201
-
Detection of HIV1 DNA in infants and children by means of the polymerase chain reaction.
Lancet. 1988 Sep 3;2(8610):538-41
PMID: 2900922
-
Detection of human papillomavirus in normal and dysplastic tissue by the polymerase chain reaction.
Lab Invest. 1988 Oct;59(4):555-9
PMID: 2845193
-
Polymerase chain reaction for detection of residual leukaemia.
Lancet. 1989 Apr 29;1(8644):928-9
PMID: 2565420
-
Detection of measles virus RNA in paraffin-embedded tissue.
Lancet. 1989 Jun 17;1(8651):1391
PMID: 2567404
-
Modification of enzymatically amplified DNA for the detection of point mutations.
Nucleic Acids Res. 1989 May 11;17(9):3606
PMID: 2726503
-
Detection of human papilloma virus in paraffin-embedded tissue using the polymerase chain reaction.
J Exp Med. 1988 Jan 1;167(1):225-30
PMID: 2826637
-
Southern blot analysis of DNA extracted from formal-saline fixed and paraffin wax embedded tissue.
J Pathol. 1988 Apr;154(4):313-20
PMID: 2838596
-
Southern blot analysis of DNA extracted from formalin-fixed pathology specimens.
Cancer Res. 1986 Jun;46(6):2964-9
PMID: 3009004
-
Purification of DNA from formaldehyde fixed and paraffin embedded human tissue.
Biochem Biophys Res Commun. 1985 Jul 16;130(1):118-26
PMID: 2992457
-
Detection of minimal residual cells carrying the t(14;18) by DNA sequence amplification.
Science. 1987 Jul 10;237(4811):175-8
PMID: 3110950
-
What price success? The continuing saga of the toxic:therapeutic ratio in the use of aminoglycoside antibiotics.
J Infect Dis. 1988 Jul;158(1):1-6
PMID: 3292658
-
Rapid detection of ras oncogenes in human tumors: applications to colon, esophageal, and gastric cancer.
Oncogene. 1989 Jul;4(7):923-8
PMID: 2666911
-
Tissue extraction of DNA and RNA and analysis by the polymerase chain reaction.
J Clin Pathol. 1990 Jun;43(6):499-504
PMID: 1696290
-
Analysis of DNA in fresh and fixed tissue by the polymerase chain reaction.
Am J Pathol. 1990 Mar;136(3):541-8
PMID: 2156429
-
Demonstration of multiple HPV types in normal cervix and in cervical squamous cell carcinoma using the polymerase chain reaction on paraffin wax embedded material.
J Clin Pathol. 1990 Jan;43(1):52-6
PMID: 2155946
-
The polymerase chain reaction. A new method of using molecular genetics for medical diagnosis.
N Engl J Med. 1990 Jan 18;322(3):178-83
PMID: 2403656
-
Rapid method for distinguishing clonal from polyclonal B cell populations in surgical biopsy specimens.
J Clin Pathol. 1990 May;43(5):429-32
PMID: 2370312
-
Analysis of DNA sequences in forty-year-old paraffin-embedded thin-tissue sections: a bridge between molecular biology and classical histology.
Cancer Res. 1988 Aug 15;48(16):4564-6
PMID: 2840197