Comparison of primers and optimization of PCR conditions for detection of Cryptosporidium parvum and Giardia lamblia in water

Article Abstract:

PCR amplification and hybridization give maximum sensitivity for the specific detection of Cryptosporidium parvum and Giardia lamblia in water samples. The optimization of primer annealing temperature and MgCl2 concentrations yields the specificities of primer pairs, which are either species or genus specific. The simultaneous detection of the two pathogenic protozoa by multiplex PCR demonstrates the amplification of 256- and 163-bp products from the 18S rRNA gene of C. parvum and heat shock protein gene of G. lamblia, respectively.

author: Stewart, Mic H., Rochelle, Paul A., De Leon, Ricardo, Wolfe, Roy L.
Giardia, Giardia lamblia

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An assay combining cell culture with reverse transcriptase PCR to detect and determine the infectivity of waterborne Crytosporidium parvum

Article Abstract:

An in vitro infectivity assay for Cryptosporidium parvum was developed by utilizing cell culture combined with reverse transcriptase-polymerase chain reaction (RT-PCR) to detect infectious organisms. The method also utilizes PCR primers that are specific for Cryptosporidium parvum to detect oocysts in environmental water samples. Furthermore, the need for multiple or nested reactions was eliminated by the spin column purification of extracted DNA from the environmental water samples.

author: Stewart, Mic H., Rochelle, Paul A., Wolfe, Roy L., Ferguson, Donna M., Handojo, Troy J., Leon, Ricardo de
Research, Methods, Analysis, Microbiology, Microbial contamination, Bacteriology

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Randomly amplified polymorphic DNA PCR analysis of bovine Cryptosporidium parvum strains isolated from the watershed of the Red River of the North

Article Abstract:

Cryptosoporidium parvum causes cryptosporidiosis and although this protozoal disease is hypothesized to be zoonotic, it has never been proven. Sixteen bovine C. parvum isolates from the Red River of the North and two bovine and two human isolates from Australia were analyzed using randomly amplified polymorphic DNA polymerase chain reaction to determine relationships among these strains analysis of Cryptosporidium. Results reveal that these isolates belong to four different strains.

author: Shianna, Kevin V., Rytter, Russell, Spanier, Jonathan G.
Pathogenic microorganisms, Protozoans, Protozoa, Pathogenic

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subjects list: Usage, Identification and classification, Polymerase chain reaction, Coccidia, Cryptosporidium
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