A rapid DNA extraction method suitable for human papillomavirus detection

Infection with oncogenic human papillomavirus (HPV) genotypes is necessary for the development of cervical cancer. Testing for HPV DNA from liquid based cervical samples can be used as an adjunct to traditional cytological screening. In addition there are ongoing viral load, genotyping, and prevalen...

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Main Authors: Brestovac, Brian, Costantino, Paul, Groth, David, Wong, M.
Format: Journal Article
Published: WILEY-LISS 2014
Subjects:
Online Access:http://hdl.handle.net/20.500.11937/26805
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author Brestovac, Brian
Costantino, Paul
Groth, David
Wong, M.
author_facet Brestovac, Brian
Costantino, Paul
Groth, David
Wong, M.
author_sort Brestovac, Brian
building Curtin Institutional Repository
collection Online Access
description Infection with oncogenic human papillomavirus (HPV) genotypes is necessary for the development of cervical cancer. Testing for HPV DNA from liquid based cervical samples can be used as an adjunct to traditional cytological screening. In addition there are ongoing viral load, genotyping, and prevalence studies. Therefore, a sensitive DNA extraction method is needed to maximize the efficiency of HPV DNA detection. The XytXtract Tissue kit is a DNA extraction kit that is rapid and so could be useful for HPV testing, particularly in screening protocols. This study was undertaken to determine the suitability of this method for HPV detection. DNA extraction from HeLa and Caski cell lines containing HPV 18 and 16 respectively together with DNA from five liquid based cervical samples were used in a HPV PCR assay. DNA was also extracted using the QIAamp DNA mini kit (Qiagen, Hilden, Germany) as a comparison. DNA extracts were serially diluted and assayed. HPV DNA was successfully detected in cell lines and cervical samples using the XytXtract Tissue kit. In addition, the XytXtract method was found to be more sensitive than the QIAmp method as determined by a dilution series of the extracted DNA. While the XytXtract method is a closed, the QIAamp method uses a spin column with possible loss of DNA through DNA binding competition of the matrix, which could impact on the final extraction efficiency. The XytXtract is a cheap, rapid and efficient method for extracting HPV DNA from both cell lines and liquid based cervical samples.
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publishDate 2014
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spelling curtin-20.500.11937-268052019-02-19T05:35:40Z A rapid DNA extraction method suitable for human papillomavirus detection Brestovac, Brian Costantino, Paul Groth, David Wong, M. XytXtract human papillomavirus liquid based cytology cervical cancer Infection with oncogenic human papillomavirus (HPV) genotypes is necessary for the development of cervical cancer. Testing for HPV DNA from liquid based cervical samples can be used as an adjunct to traditional cytological screening. In addition there are ongoing viral load, genotyping, and prevalence studies. Therefore, a sensitive DNA extraction method is needed to maximize the efficiency of HPV DNA detection. The XytXtract Tissue kit is a DNA extraction kit that is rapid and so could be useful for HPV testing, particularly in screening protocols. This study was undertaken to determine the suitability of this method for HPV detection. DNA extraction from HeLa and Caski cell lines containing HPV 18 and 16 respectively together with DNA from five liquid based cervical samples were used in a HPV PCR assay. DNA was also extracted using the QIAamp DNA mini kit (Qiagen, Hilden, Germany) as a comparison. DNA extracts were serially diluted and assayed. HPV DNA was successfully detected in cell lines and cervical samples using the XytXtract Tissue kit. In addition, the XytXtract method was found to be more sensitive than the QIAmp method as determined by a dilution series of the extracted DNA. While the XytXtract method is a closed, the QIAamp method uses a spin column with possible loss of DNA through DNA binding competition of the matrix, which could impact on the final extraction efficiency. The XytXtract is a cheap, rapid and efficient method for extracting HPV DNA from both cell lines and liquid based cervical samples. 2014 Journal Article http://hdl.handle.net/20.500.11937/26805 10.1002/jmv.23882 WILEY-LISS fulltext
spellingShingle XytXtract
human papillomavirus
liquid based cytology
cervical cancer
Brestovac, Brian
Costantino, Paul
Groth, David
Wong, M.
A rapid DNA extraction method suitable for human papillomavirus detection
title A rapid DNA extraction method suitable for human papillomavirus detection
title_full A rapid DNA extraction method suitable for human papillomavirus detection
title_fullStr A rapid DNA extraction method suitable for human papillomavirus detection
title_full_unstemmed A rapid DNA extraction method suitable for human papillomavirus detection
title_short A rapid DNA extraction method suitable for human papillomavirus detection
title_sort rapid dna extraction method suitable for human papillomavirus detection
topic XytXtract
human papillomavirus
liquid based cytology
cervical cancer
url http://hdl.handle.net/20.500.11937/26805