Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Search research articles

Related Experiment Videos

A highly sensitive nonisotopic detection method for in situ hybridization.

¹Department of Pathology, Norwegian Radium Hospital, Montebello, Oslo, Norway. Ruth.holm@labmed.uio.no

Applied Immunohistochemistry & Molecular Morphology : AIMM

|August 11, 2000

Summary

Comparing nonisotopic detection systems for in situ hybridization, the APAAP method demonstrated superior sensitivity for detecting human papillomavirus (HPV) 16 DNA in formalin-fixed tissues, even at single-copy levels.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by

Same author

Corrigendum to 'Is Oral Absorption of Vigabatrin Carrier-mediated?' [European Journal of Pharmaceutical Sciences 69 (2015) 10-18].

European journal of pharmaceutical sciences : official journal of the European Federation for Pharmaceutical Sciences·2021

Same author

Fasted and fed state human duodenal fluids: Characterization, drug solubility, and comparison to simulated fluids and with human bioavailability.

European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V·2021

Same author

Six years of progress in the oral biopharmaceutics area - A summary from the IMI OrBiTo project.

European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V·2020

Same author

Solution or suspension - Does it matter for lipid based systems? In vivo studies of chase dosing lipid vehicles with aqueous suspensions of a poorly soluble drug.

European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V·2017

Same author

A survey on IVIVC/IVIVR development in the pharmaceutical industry - Past experience and current perspectives.

European journal of pharmaceutical sciences : official journal of the European Federation for Pharmaceutical Sciences·2017

Same author

Is oral absorption of vigabatrin carrier-mediated?

European journal of pharmaceutical sciences : official journal of the European Federation for Pharmaceutical Sciences·2015

Area of Science:

Molecular Biology
Pathology
Virology

Background:

In situ hybridization (ISH) detects specific DNA/RNA in tissues.
Nonisotopic ISH offers speed and precise localization but often lacks sensitivity.
Sensitivity in nonisotopic ISH is highly dependent on the chosen detection system.

Purpose of the Study:

To evaluate and compare the sensitivity of three nonisotopic detection systems for in situ hybridization.
To identify the most sensitive method for detecting human papillomavirus (HPV) 16 DNA in challenging samples.

Main Methods:

Compared STAV-FITC, APAAP, and tyramide signal amplification (TSA) detection systems.
Utilized biotin-labeled HPV 16 probes on HPV16-positive cell lines (CaSki, SiHa) and cervical carcinomas.

Related Experiment Videos

Employed formalin-fixed, paraffin-embedded tissues and cells for all methods.

Main Results:

All three methods provided satisfactory staining in high-HPV16 copy samples (CaSki cells, carcinomas).
Only the APAAP method successfully detected HPV16 DNA in SiHa cells with low viral copy numbers (1-2 per cell).
APAAP method showed significantly higher sensitivity compared to STAV-FITC and TSA methods.

Conclusions:

The APAAP method offers the highest sensitivity among the evaluated nonisotopic detection systems for ISH.
APAAP is capable of detecting single viral DNA copies in formalin-fixed, paraffin-embedded cervical cancer specimens.
This finding is crucial for sensitive molecular diagnostics in pathology.