+ Resolve Article
+ Follow Us
Follow on FacebookFollow on Facebook
Follow on TwitterFollow on Twitter

+ Translate
+ Subscribe to Site Feed
GeoScience Most Shared ContentMost Shared Content

Identification of Cryptosporidium spp. oocysts in United Kingdom noncarbonated natural mineral waters and drinking waters by using a modified nested PCR-restriction fragment length polymorphism assay

, : Identification of Cryptosporidium spp. oocysts in United Kingdom noncarbonated natural mineral waters and drinking waters by using a modified nested PCR-restriction fragment length polymorphism assay. Applied and Environmental Microbiology 69(7): 4183-4189

We describe a nested PCR-restriction fragment length polymorphism (RFLP) method for detecting low densities of Cryptosporidium spp. oocysts in natural mineral waters and drinking waters. Oocysts were recovered from seeded 1-liter volumes of mineral water by filtration through polycarbonate membranes and from drinking waters by filtration, immunomagnetizable separation, and filter entrapment, followed by direct extraction of DNA. The DNA was released from polycarbonate filter-entrapped oocysts by disruption in lysis buffer by using 15 cycles of freeze-thawing (1 min in liquid nitrogen and 1 min at 65 degrees C), followed by proteinase K digestion. Amplicons were readily detected from two to five intact oocysts on ethidium bromide-stained gels. DNA extracted from Cryptosporidium parvum oocysts, C. muris (RN 66), C. baileyi (Belgium strain, LB 19), human-derived C. meleagridis, C. felis (DNA from oocysts isolated from a cat), and C. andersoni was used to demonstrate species identity by PCR-RFLP after simultaneous digestion with the restriction enzymes DraI and VspI. Discrimination between C. andersoni and C. muris isolates was confirmed by a separate, subsequent digestion with DdeI. Of 14 drinking water samples tested, 12 were found to be positive by microscopy, 8 were found to be positive by direct PCR, and 14 were found to be positive by using a nested PCR. The Cryptosporidium species detected in these finished water samples was C. parvum genotype 1. This method consistently and routinely detected >5 oocysts per sample.


PMID: 12839797

DOI: 10.1128/AEM.69.7.4183-4189.2003

Other references

Ma, X.; Ge, J.; Zhang, J-ying.; Wen, W., 2004: Survey of behaviors and knowledge about HIV/AIDS among intravenous drug users at a city in Sichuan Province. To understand high risk behaviors related to HIV/AIDS and the intravenous drug users' knowledge about HIV/AIDS. Forty-eight intravenous drug users (IDUs) were interviewed by using questionnaires. The survey showed that 75% of the subjects wer...

Radzyukevich, T.L., 1995: Reinnervation of mixed muscle from the frog Rana temporaria by regenerating homogeneous nerve. Mixed muscle m. iliofibularis from the frog Rana temporaria, consisting of monosynaptically innervated phasic and polysynaptically innervated postural muscle fibers, was reinnervated by homogeneous tailor's nerve having no axons of tonic moto...

Popovici, Z., 1982: Jejunal invagination after antrectomy and gastrostomy in postcaustic esophagogastric stenoses. The author presents 3 cases of jejunal invagination in a lot of 120 cases of post-caustic oeso-gastric stenoses that were operated. In two of the patients the invagination developed following antrectomy and in one case it occurred after gastrostom...

Rawle, F.C.; Knowles, B.B.; Ricciardi, R.P.; Brahmacheri, V.; Duerksen-Hughes, P.; Wold, W.S.; Gooding, L.R., 1991: Specificity of the mouse cytotoxic T lymphocyte response to adenovirus 5. E1A is immunodominant in H-2b, but not in H-2d or H-2k mice. The Ag specificity and MHC restriction of the CTL response to adenovirus 5 (Ad5) in three strains of mice, C57BL/10 (H-2b), BALB/c (H-2d), and C3H/HeJ (H-2k), were tested. Polyclonal Ad5-specific CTL were prepared by priming mice in vivo with live...

Ashwell M.; Jennings G.; Stirling D.M.; Trayhurn P., 1986: Decrease in mitochondrial gdp binding without parallel changes in uncoupling protein in brown adipose tissue of the guinea pig. Biochemical Society Transactions 14(2): 283-284

Johnston, C.S.; Yen, M.F., 1994: Vitamin C alters insulin response to a glucose challenge in normoglycemic adults. FASEB Journal 8(4-5): A446

Sollewijn Gelpke, M.D.; Sheng, D.; Gold, M.H., 2000: MnII is not a productive substrate for wild-type or recombinant lignin peroxidase isozyme H2. The glyceraldehyde-3-phosphate dehydrogenase (gpd) gene promoter was used to drive the homologous expression of the lignin peroxidase (LiP) isozyme H2 gene in primary metabolic cultures of Phanerochaete chrysosporium. The molecular mass, pI, and o...

Okai H.; Ono K.; Kanehisa H., 1972: Relationship between the taste and chemical structure of alanine containing peptide. Seikagaku 44(11): 947

E.Denshary E.S.M., 1972: Influence of the concomitant administration of tolbutamide and chlorpromazine on certain adrenal functions in rat. Egyptian Journal Of Pharmaceutical Sciences: 301-307

Hrabar S.V.; Potter Paul Edwin, 1970: Cross-bedding in the Tanglewood Limestone member of the Lexington limestone (Ordovician), central Kentucky. Abstracts with Programs Geological Society of America: Pages 218-219. 1970.