If you are experiencing problems downloading PDF or HTML fulltext, our helpdesk recommend clearing your browser cache and trying again. If you need help in clearing your cache, please click here . Still need help? Email help@ingentaconnect.com

DNA extraction and PCR assays for detection of Toxoplasma gondii

$48.00 plus tax (Refund Policy)

Download / Buy Article:

Abstract:

Edvisson B, Jalal S, Nord CE, Pedersen BS, Evengård B. DNA extraction and PCR assays for detection of Toxoplasma gondii. APMIS 2004;112:342–8.

For detection of Toxoplasma gondii we compared the sensitivity of two different DNA extraction methods and three different PCR assays. Sensitivities of DNA extraction by QIAamp DNA mini Kit or MagNa pure followed by PCR, nested PCR and oligochromatography or Light Cycler PCR using either SYBR green chemistry or TaqMan probe were compared. No significant difference between extraction methods was found using pure T. gondii tachyzoites. Spiked blood samples, 104 to 10 parasites per sample, generated no difference in sensitivity between the two DNA extraction methods when analysed by nested PCR detected by oligochromatography or analysed by Light Cycler PCR TaqMan. In spiked blood samples Light Cycler PCR SYBR green was unable to detect the parasite and a reduction in sensitivity was observed with the TaqMan assay. Conventional PCR was more sensitive when DNA was extracted from the spiked samples using the QIAamp DNA mini Kit. Conventional and nested PCR were found to be more sensitive than Light Cycler PCR TaqMan using the QIAamp DNA mini Kit. It was not possible to use Light Cycler PCR SYBR green in blood samples. Conventional PCR was more sensitive for detection of T. gondii in spiked blood samples using QIAamp DNA mini Kit DNA extraction, suggesting that the choice of DNA extraction method may affect PCR assays differently.

Keywords: DNA extraction; PCR; TaqMan; Toxoplasma gondii; oligochromatography; spiked blood samples

Document Type: Research Article

DOI: http://dx.doi.org/10.1111/j.1600-0463.2004.apm1120604.x

Affiliations: 1: Department Laboratory Medicine, F82, Karolinska Institute, Huddinge University hospital, Stockholm, Sweden 2: Department Gynecology and Obstetrics, Rikshospitalet, University of Oslo, Norway

Publication date: June 1, 2004

Related content

Tools

Favourites

Share Content

Access Key

Free Content
Free content
New Content
New content
Open Access Content
Open access content
Subscribed Content
Subscribed content
Free Trial Content
Free trial content
Cookie Policy
X
Cookie Policy
ingentaconnect website makes use of cookies so as to keep track of data that you have filled in. I am Happy with this Find out more