Detection of Cryptosporidium parvum in raw milk by PCR and oligonucleotide probe hybridization
| Autor: | I Laberge, A Ibrahim, Mansel W. Griffiths, J R Barta |
|---|---|
| Rok vydání: | 1996 |
| Předmět: |
animal diseases
Molecular Sequence Data Polymerase Chain Reaction Sensitivity and Specificity Applied Microbiology and Biotechnology Microbiology law.invention law parasitic diseases Animals Humans Polymerase chain reaction Cryptosporidium parvum Base Sequence Ecology biology Nucleic Acid Hybridization Cryptosporidium DNA Protozoan Raw milk biology.organism_classification Molecular biology Eimeria acervulina Milk Protozoa Cattle Primer (molecular biology) Oligonucleotide Probes Oligomer restriction Research Article Food Science Biotechnology |
| Zdroj: | Applied and Environmental Microbiology. 62:3259-3264 |
| ISSN: | 1098-5336 0099-2240 |
| DOI: | 10.1128/aem.62.9.3259-3264.1996 |
| Popis: | Cryptosporidium spp. are potential contaminants of food. Suspected cases of food-borne cryptosporidiosis are rarely confirmed because of the limited numbers of oocysts in the samples and the lack of sensitive detection methods adaptable to food. PCR was investigated as a means of overcoming this problem. A PCR assay was designed for the specific amplification of a previously sequenced portion of an oocyst protein gene fragment of Cryptosporidium parvum (N. C. Lally, G. D. Baird, S. J. McQuay, F. Wright, and J. J. Oliver, Mol. Biochem. Parasitol. 56:69-78, 1992) and compared with the primer set of Laxer et al. (M. A. Laxer, B. K. Timblin, and R. J. Patel, Am. J. Trop. Med. Hyg. 45:688-694, 1991). The PCR products were hybridized with digoxigenin-labeled internal probes and detected by chemiluminescence to enhance sensitivity. The two sets of primers were compared with regard to their sensitivity and specificity by using a variety of human and animal isolates of C. parvum and related parasites. Both assays enabled the detection of 1 to 10 oocysts in 20 ml of artificially contaminated raw milk. The assay based on the PCR set and probe of Laxer et al. detected DNAs from Eimeria acervulina and Giardia intestinalis. The new assay has good specificity for C. parvum bovine isolates and hence has a better potential for monitoring the prevalence of C. parvum in raw milk and other environmental samples. |
| Databáze: | OpenAIRE |
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