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International Journal of Environment Science and Technology
Center for Environment and Energy Research and Studies (CEERS)
ISSN: 1735-1472
EISSN: 1735-2630
Vol. 4, No. 2, 2007, pp. 241-246
Bioline Code: st07031
Full paper language: English
Document type: Research Article
Document available free of charge

International Journal of Environment Science and Technology, Vol. 4, No. 2, 2007, pp. 241-246

 en Evaluation of QIAamp DNA mini kit for removing of inhibitors in detection of Cryptosporidium parvum check for this species in other resources oocysts in water samples by a nested– PCR assay
Nikaeen, M. & Makimura, K.

Abstract

In recent years, there has been a dramatic increase in the occurrence of waterborne disease outbreaks caused by the Cryptosporidium parvum check for this species in other resources , and presence of this protozoan parasite in drinking water is a significant health problem faced by the water industry. A new strategy for detection of Cryptosporidium oocysts in water samples is PCR– based techniques. In this study a nested– PCR assay was designed for the specific amplification of a 199 bp DNA fragment of the gene encoding the heat shock protein (hsp70) of Cryptosporidium parvum oocysts. In order to prevent the inhibition of PCR amplification by substances contained in water samples, three DNA purification methods including QIAamp DNA mini kit, InstaGene Matrix, MagExtractor – Genome were compared in concentrates of tap water samples spiked with the oocysts. After it was found that the QIAamp is only efficient purification technique, the efficiency of QIAamp and immunomagnetic separation for nested–PCR assay of various water samples was compared. The results show that QIAamp provide a useful and rapid tool for removing of PCR inhibitors. It seems that QIAamp purification-nested PCR assay is a sensitive, rapid and cost effective method for detection of Cryptosporidium parvum oocysts in clean water samples with turbidity < 2 nephelometric turbidity unit (NTU).

Keywords
Cryptosporidium, PCR, detection, QIAamp, water, purification

 
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