A fast and sensitive nucleic acid extraction method for the detection of Cryptosporidium by PCR in environmental water samples

被引：2

作者：

Dellundé, J ^{[1
]}

Pina, S ^{[1
]}

Jofre, J ^{[1
]}

Lucena, F ^{[1
]}

机构：

[1] Univ Barcelona, Dept Microbiol, Barcelona, Spain

来源：

2ND WORLD WATER CONGRESS: WATER AND HEALTH-MICROBIOLOGY, MONITORING AND DISINFECTION | 2002年 / 2卷 / 03期

关键词：

cryptosporidium; environmental; excystation; PCR;

D O I：

10.2166/ws.2002.0090

中图分类号：

X [环境科学、安全科学];

学科分类号：

08 ; 0830 ;

摘要：

A new protocol based on a combination of an excystation process followed by a nucleic acid extraction that combines the lysing and nuclease-inactivating properties of the chaotropic agent guanidinium thiocyanate and the nucleic acid-binding properties of silica particles is described for the extraction and purification of nucleic acids from Cryptosporidium. The application of nested and/or semi-nested PCR using different external and internal primers to DNA extracted by this method from seeded and naturally occurring Cryptosporidium oocysts concentrated and purified from environmental samples detects numbers of oocysts ranging from 20 to 50. The method is feasible, detects mostly excystable oocysts and no problems of inhibition of PCR were observed when applied to environmental samples.

引用

页码：95 / 100

页数：6

共 11 条

[1] RAPID AND SIMPLE METHOD FOR PURIFICATION OF NUCLEIC-ACIDS [J].