EurekaMag.com logo
+ Site Statistics
References:
53,869,633
Abstracts:
29,686,251
+ Search Articles
+ Subscribe to Site Feeds
EurekaMag Most Shared ContentMost Shared
EurekaMag PDF Full Text ContentPDF Full Text
+ PDF Full Text
Request PDF Full TextRequest PDF Full Text
+ Follow Us
Follow on FacebookFollow on Facebook
Follow on TwitterFollow on Twitter
Follow on LinkedInFollow on LinkedIn

+ Translate

Comparison of most probable number-PCR and most probable number-foci detection method for quantifying infectious Cryptosporidium parvum oocysts in environmental samples



Comparison of most probable number-PCR and most probable number-foci detection method for quantifying infectious Cryptosporidium parvum oocysts in environmental samples



Journal of Microbiological Methods 67(2): 363-372



Microbial contamination of public water supplies is of significant concern, as numerous outbreaks, including Cryptosporidium, have been reported worldwide. Detection and enumeration of Cryptosporidium parvum oocysts in water supplies is important for the prevention of future cryptosporidiosis outbreaks. In addition to not identifying the oocyst species, the U.S. EPA Method 1622 does not provide information on oocyst viability or infectivity. As such, current detection strategies have been coupled with in vitro culture methods to assess oocyst infectivity. In this study, a most probable number (MPN) method was coupled with PCR (MPN-PCR) to quantify the number of infectious oocysts recovered from seeded raw water concentrates. The frequency of positive MPN-PCR results decreased as the oocyst numbers decreased. Similar results were observed when MPN was coupled to the foci detection method (MPN-FDM), which was done for comparison. For both methods, infectious oocysts were not detected below 103 seeded oocysts and the MPN-PCR and MPN-FDM estimates for each seed dose were generally within one-log unit of directly enumerated foci of infection. MPN-PCR estimates were 0.25, 0.54, 0 and 0.66 log10 units higher than MPN-FDM estimates for the positive control, 105, 104 and 103 seed doses, respectively. The results show the MPN-PCR was the better method for the detection of infectious C. parvum oocysts in environmental water samples.

(PDF emailed within 0-6 h: $19.90)

Accession: 011884741

Download citation: RISBibTeXText

PMID: 16730821

DOI: 10.1016/j.mimet.2006.04.007



Related references

A most-probable-number assay for enumeration of infectious Cryptosporidium parvum oocysts. Applied and Environmental Microbiology 65(9): 3936-3941, 1999

Detection of Cryptosporidium parvum in secondary effluents using a most probable number-polymerase chain reaction assay. Water Environment Research 75(4): 292-299, 2003

Detection of infectious Cryptosporidium parvum oocysts in environmental water samples using an integrated cell culture-PCR (CC-PCR) system. Water, Air, and Soil Pollution 123(1/4): 53-65, 2000

Seasonal change in the number of Cryptosporidium parvum oocysts in water samples from the rivers in Hokkaido, Japan, detected by the ferric sulfate flocculation method. Journal of Veterinary Medical Science 65(1): 121-123, 2003

A comparison of filter capture and precultivation for quantitative detection of infectious hematopoietic necrosis virus by using polymerase chain reaction combined with the most probable number method. Journal of Aquatic Animal Health 11(2): 154-157, June, 1999

Evaluation of immunomagnetic separation for recovery of infectious Cryptosporidium parvum oocysts from environmental samples. Applied and Environmental Microbiology 65(2): 841-845, 1999

A comparison of filter capture and precultivation for quantitative detection of infectious hematopoietic necrosis virus (IHNV) by using polymerase chain reaction (PCR) combined with the most probable number (MPN) method. Journal of Aquatic Animal Health 11(2): 154-157, 1999

Modified most probable number technique for the specific determination of escherichia coli from environmental samples using a fluorogenic method. Journal of Microbiological Methods 12(3-4): 235-246, 1990

Detection of infectious Cryptosporidium parvum oocysts in surface and filter backwash water samples by immunomagnetic separation and integrated cell culture-PCR. Applied and Environmental Microbiology 65(8): 3427-3432, 1999

Detection of cryptosporidia and Cryptosporidium parvum oocysts in environmental water samples by immunomagnetic separation-polymerase chain reaction. Journal of Applied Microbiology 89(1): 5-10, 2000

Comparison of immunofluorescence assay and immunomagnetic electrochemiluminescence in detection of Cryptosporidium parvum oocysts in karst water samples. Journal of microbiological methods 53(1): 17-26, 2003

Detection of a small number of Cryptosporidium parvum oocysts by sugar floatation and sugar centrifugation methods. Journal of Veterinary Medical Science 68(11): 1191-1193, 2006