Dataset Information

Rapid and sensitive detection of single cryptosporidium oocysts from archived glass slides.

ABSTRACT: In this study we report on the development and application of a novel method for efficiently extracting and detecting single Cryptosporidium oocysts from archived glass slides. Laser capture microscopy was used to extract low numbers of oocysts from archived glass slides. Highly sensitive real-time PCR methods were then developed to enable the rapid detection and identification of Cryptosporidium oocysts from these samples. The method was applied to fecal smears stained with a variety of standard oocyst stains and water samples. This application, with samples derived from both public health and water service laboratories, highlighted the strong potential of this method to be used as a rapid high-throughput screening tool for the routine monitoring of Cryptosporidium and other medically important pathogens from clinical, veterinary, and environmental water samples. Importantly, the application of our protocol could be used to type Cryptosporidium and other pathogens from stored archived glass slides in public health and water service laboratories, providing vital epidemiological updates and helping to identify and trace pathogens and their routes of infection and ultimately improve their control.

SUBMITTER: Sunnotel O

PROVIDER: S-EPMC1594691 | biostudies-literature | 2006 Sep

REPOSITORIES: biostudies-literature

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Rapid and sensitive detection of single cryptosporidium oocysts from archived glass slides.

Sunnotel O O Snelling W J WJ Xiao L L Moule K K Moore J E JE Millar B Cherie BC Dooley J S G JS Lowery C J CJ

Journal of clinical microbiology 20060901 9

In this study we report on the development and application of a novel method for efficiently extracting and detecting single Cryptosporidium oocysts from archived glass slides. Laser capture microscopy was used to extract low numbers of oocysts from archived glass slides. Highly sensitive real-time PCR methods were then developed to enable the rapid detection and identification of Cryptosporidium oocysts from these samples. The method was applied to fecal smears stained with a variety of standar ...[more]

PMID: 16954262

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Project description:This study investigated the use of biofilms to monitor Cryptosporidium in water. Benthic rock and submersible slide biofilms were sampled upstream and downstream of point sources in a suburban watershed in southeastern Pennsylvania. More oocysts were detected in biofilms scraped from rocks downstream than upstream of a wastewater treatment plant (WWTP) (19 versus 5, respectively; n = 1). Although not statistically significant, Cryptosporidium oocysts were detected more frequently, and in greater numbers, in biofilms grown on slides downstream than upstream of this same WWTP (83.3% positive samples [n = 12] versus 45.5% positive samples [n = 11], respectively; P = 0.0567). Similarly, Cryptosporidium oocysts were detected more frequently, and in greater numbers, in rock biofilms collected downstream than upstream of a stormwater outfall impacted by defective sewer laterals (50% positive samples downstream and 17% positive samples upstream; n = 6; P = 0.2207). While oocyst detection data obtained by slide biofilms versus filters did not necessarily agree on a given day, there was no seasonal difference in the frequency of oocyst detection (P > 0.05) or numbers of oocysts detected (P > 0.05) whether the water was monitored by filtration or slide biofilm sampling. Within any given season, there was no difference in the frequency of oocyst detection (P > 0.05) or the numbers of oocysts detected (P > 0.05) whether the water was monitored by filtration or slide biofilm sampling. These data show that oocyst detection in biofilms is comparable to oocyst detection in filtered water samples. Biofilm sampling offers significant cost savings compared to the filtration-based EPA Method 1623.1 and could be used to identify watershed locations at potential risk for increased oocyst loads.IMPORTANCE Monitoring Cryptosporidium occurrence in watersheds that provide drinking water is necessary to determine where limited resources should most effectively be directed to protect consumers from waterborne exposure to pathogenic oocysts. Biofilms are a useful tool to monitor complex watersheds and identify point sources of Cryptosporidium oocyst contamination that need to be managed to protect public health. Compared to EPA Method 1623.1, the cost benefit of using biofilms to monitor for Cryptosporidium contamination will enable utilities to sample water supplies more frequently, and at more locations, than is currently possible given limited operating budgets. Biofilm sampling could be used to identify high-risk regions within a large, complex watershed and the associated water treatment plants at potential risk for increased oocyst loads in the water supply; this information could then be used to select the locations within the watershed where the more expensive EPA Method 1623.1 is warranted.

Dataset Information

Rapid and sensitive detection of single cryptosporidium oocysts from archived glass slides.

Publications

Rapid and sensitive detection of single cryptosporidium oocysts from archived glass slides.

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