Literature DB >> 2844861

Non-isotopic detection of in situ nucleic acid in cervix: an updated protocol.

J Burns1, A K Graham, J O McGee.   

Abstract

Recently, sensitive non-isotopic in situ hybridisation (NISH) methodology for the detection of human DNA and human papilloma virus (HPV) DNA in archival paraffin blocks of cervix was described. An amended protocol, now used in this laboratory for detection of these genes by NISH is presented. The amendments include the following: protease digestion at 37 degrees C; tissue dehydration in air rather than ethanol; stringency washing in formamide solution; blocking non-specific binding of avidin alkaline phosphatase with a modified buffer; and increasing the concentration of avidin alkaline phosphatase for detecting low abundance DNA. These changes simplify and increase the sensitivity of the protocol such that "Y" chromosome repeats are visualised in almost all female cells.

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Year:  1988        PMID： 2844861      PMCID： PMC1141624          DOI： 10.1136/jcp.41.8.897

Source DB:  PubMed          Journal:  J Clin Pathol        ISSN： 0021-9746            Impact factor:   3.411


  4 in total

1.  Detection of low copy human papilloma virus DNA and mRNA in routine paraffin sections of cervix by non-isotopic in situ hybridisation.

Authors:  J Burns; A K Graham; C Frank; K A Fleming; M F Evans; J O McGee
Journal:  J Clin Pathol       Date:  1987-08       Impact factor: 3.411

2.  Human papillomavirus type 13 and focal epithelial hyperplasia of the oral mucosa: DNA hybridization on paraffin-embedded specimens.

Authors:  R P Henke; K Milde-Langosch; T Löning; H S Koppang
Journal:  Virchows Arch A Pathol Anat Histopathol       Date:  1987

3.  Detection of sub-picogram quantities of specific DNA sequences on blot hybridization with biotinylated probes.

Authors:  V T Chan; K A Fleming; J O McGee
Journal:  Nucleic Acids Res       Date:  1985-11-25       Impact factor: 16.971

4.  Chromosomal localization of genes by scanning electron microscopy using in situ hybridization with biotinylated probes: Y chromosome repetitive sequences.

Authors:  D J Ferguson; J Burns; D Harrison; J A Jonasson; J O McGee
Journal:  Histochem J       Date:  1986-05
  4 in total
  11 in total

Review 1.  In situ hybridisation in perspective.

Authors:  A Warford; I Lauder
Journal:  J Clin Pathol       Date:  1991-03       Impact factor: 3.411

2.  Simultaneous in situ genotyping and phenotyping of human papillomavirus cervical lesions: comparative sensitivity and specificity.

Authors:  A K Graham; C S Herrington; O McGee J
Journal:  J Clin Pathol       Date:  1991-02       Impact factor: 3.411

3.  Discrimination of closely homologous HPV types by nonisotopic in situ hybridization: definition and derivation of tissue melting temperatures.

Authors:  C S Herrington; A K Graham; D M Flannery; J Burns; J O McGee
Journal:  Histochem J       Date:  1990-10

4.  Human papillomaviruses in anogenital warts in children: typing by in situ hybridisation.

Authors:  A F Padel; V A Venning; M F Evans; A M Quantrill; K A Fleming
Journal:  BMJ       Date:  1990-06-09

5.  Interphase cytogenetics using biotin and digoxigenin labelled probes I: relative sensitivity of both reporter molecules for detection of HPV16 in CaSki cells.

Authors:  C S Herrington; J Burns; A K Graham; M Evans; J O McGee
Journal:  J Clin Pathol       Date:  1989-06       Impact factor: 3.411

6.  Interphase cytogenetics using biotin and digoxigenin labelled probes II: Simultaneous differential detection of human and papilloma virus nucleic acids in individual nuclei.

Authors:  C S Herrington; J Burns; A K Graham; B Bhatt; J O McGee
Journal:  J Clin Pathol       Date:  1989-06       Impact factor: 3.411

7.  Discrimination of closely homologous human genomic and viral sequences in cells and tissues: further characterization of Tmt.

Authors:  C S Herrington; J O McGee
Journal:  Histochem J       Date:  1994-07

8.  Comparative analysis of human papillomavirus detection by dot blot hybridisation and non-isotopic in situ hybridisation.

Authors:  G Troncone; S M Anderson; C S Herrington; M L de Angelis; H Noell; J A Chimera; J O'D McGee
Journal:  J Clin Pathol       Date:  1992-10       Impact factor: 3.411

9.  Chromosome in situ hybridization on formalin-fixed mammary tissue using non-isotopic, non-fluorescent probes: technical considerations and biological implications.

Authors:  K Dhingra; A Sahin; J Supak; S Y Kim; G Hortobagyi; W N Hittelman
Journal:  Breast Cancer Res Treat       Date:  1992       Impact factor: 4.872

10.  Detection of human papillomavirus in matched cervical smears and biopsy specimens by non-isotopic in situ hybridisation.

Authors:  G Troncone; C S Herrington; K Cooper; M L de Angelis; J O McGee
Journal:  J Clin Pathol       Date:  1992-04       Impact factor: 3.411
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