Comparison of five DNA extraction methods and optimization of a b1 gene nested PCR (nPCR) for detection of Toxoplasma gondii tissue cyst in mouse brain.
Southeast Asian J Trop Med Public Health
;
2005 Nov; 36(6): 1377-82
Article
in English
| IMSEAR
| ID: sea-31491
ABSTRACT
Toxoplasmosis, caused by Toxoplasma gondii, is an important parasitic disease worldwide. Different techniques have been developed for T. gondii detection. At present, polymerase chain reaction (PCR) has been widely used. However, PCR for identifying T. gondii remains unsatisfactory in many laboratories because of lack of standardization and variations in efficiency. In the present study, we optimized a nested PCR protocol (n-PCR) in order to compare the amplification of T. gondii DNA, after being extracted from mouse brain by five different DNA extraction methods including phenol chloroform, QIAamp DNA minikit, Genomic DNA purification kit and Chelex with or without proteinase K. All DNA extraction methods were able to extract DNA from a single tissue cyst from mouse brain. However, among the five DNA extraction methods, the Chelex without proteinase K appeared to be the most rapid and easiest.
Full text:
Available
Index:
IMSEAR (South-East Asia)
Main subject:
Toxoplasma
/
Brain
/
Female
/
Male
/
Toxoplasmosis, Animal
/
Polymerase Chain Reaction
/
DNA, Protozoan
/
Genes, Protozoan
/
DNA Primers
/
Cysts
Type of study:
Diagnostic study
Language:
English
Journal:
Southeast Asian J Trop Med Public Health
Year:
2005
Type:
Article
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