[Preparation of DNA from Cryptosporidium parvum oocysts for PCR detection].
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi
; 23(4): 228-30, 2005 Aug 30.
Article
em Zh
| MEDLINE
| ID: mdl-16296614
ABSTRACT
OBJECTIVE:
To establish three methods of DNA extraction from Cryptosporidium parvum oocysts and test by PCR.METHODS:
After three freeze-thaw cycles, three kinds of templates were extracted from the oocysts by Chelex-100, phenol/chloroform or genomic DNA purification system kit, and used for PCR detection. According to the sequence of a C. parrum gene (L.16996), a pair of primers was designed and synthesized, and used for PCR. The sensitivity of the template by Chelex 100 method was also tested by PCR.RESULTS:
One 446 bp PCR product was observed by agarose gel electrophoresis for all three kinds of templates. The PCR sensitivity by Chelex-100 extracted DNA reached for detection of a specimen containing only 1/2 oocyst.CONCLUSION:
The three kinds of extraction can all be served as templates for PCR detection of C. parvum oocysts, while Chelex 100 method is simpler, quicker and more reliable for DNA extraction of the parasite.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Reação em Cadeia da Polimerase
/
DNA de Protozoário
/
Cryptosporidium parvum
/
Oocistos
Tipo de estudo:
Diagnostic_studies
Limite:
Animals
/
Child
/
Humans
Idioma:
Zh
Ano de publicação:
2005
Tipo de documento:
Article