Last data update: Jun 24, 2024. (Total: 47078 publications since 2009)
Records 1-4 (of 4 Records) |
Query Trace: Mull BJ [original query] |
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Real-time PCR and Sequencing Assays for Rapid Detection and Identification of Avian Schistosomes in Environmental Samples.
Narayanan J , Mull BJ , Brant SV , Loker ES , Collinson J , Secor WE , Hill VR . Appl Environ Microbiol 2015 81 (12) 4207-15 ![]() ![]() Cercarial dermatitis, also known as swimmer's itch, is an allergenic skin reaction followed by intense itching caused by schistosome cercariae penetrating human skin. Cercarial dermatitis outbreaks occur globally, and are frequently associated with fresh water lakes and occasionally with marine or estuarine waters where year-round or migratory birds reside. In this study, a broadly reactive TaqMan assay was developed targeting 18S ribosomal RNA (rDNA) gene sequences based on a genetically diverse panel of schistosome isolates representing 13 genera and 20 species. A PCR assay was also developed to amplify a 28S ribosomal RNA (rDNA) gene region for subsequent sequencing to identify schistosomes. When applied to surface water samples seeded with Schistosoma mansoni cercariae, the 18S TaqMan assay enabled detection at a level of 5 S. mansoni cercariae in 100 L of lake water. The 18S TaqMan and 28S PCR-sequencing assays were also applied to 100-L water samples collected from lakes in Nebraska and Wisconsin where there were reported dermatitis outbreaks. Avian schistosome DNA was detected in 11 of 34 lake water samples using the TaqMan assay. Further 28S sequence analysis of positive samples confirmed the presence, and provided preliminary identification of avian schistosomes in ten of the 11 samples. These data indicate that the broadly schistosome-reactive TaqMan assay can be effective for rapid screening of large-volume water samples for detection of avian schistosomes, thereby facilitating timely response actions to mitigate or prevent dermatitis outbreaks. Additionally, samples positive by the 18S TaqMan assay can be further assayed using the 28S sequencing assay to both to confirm the presence of schistosomes and contribute to their identification. |
Comparison of real-time PCR methods for the detection of Naegleria fowleri in surface water and sediment
Streby A , Mull BJ , Levy K , Hill VR . Parasitol Res 2015 114 (5) 1739-46 ![]() Naegleria fowleri is a thermophilic free-living ameba found in freshwater environments worldwide. It is the cause of a rare but potentially fatal disease in humans known as primary amebic meningoencephalitis. Established N. fowleri detection methods rely on conventional culture techniques and morphological examination followed by molecular testing. Multiple alternative real-time PCR assays have been published for rapid detection of Naegleria spp. and N. fowleri. Foursuch assays were evaluated for the detection of N. fowleri from surface water and sediment. The assays were compared for thermodynamic stability, analytical sensitivity and specificity, detection limits, humic acid inhibition effects, and performance with seeded environmental matrices. Twenty-one ameba isolates were included in the DNA panel used for analytical sensitivity and specificity analyses. N. fowleri genotypes I and III were used for method performance testing. Two of the real-time PCR assays were determined to yield similar performance data for specificity and sensitivity for detecting N. fowleri in environmental matrices. |
Environmental surveillance for toxigenic Vibrio cholerae in surface waters of Haiti.
Kahler AM , Haley BJ , Chen A , Mull BJ , Tarr CL , Turnsek M , Katz LS , Humphrys MS , Derado G , Freeman N , Boncy J , Colwell RR , Huq A , Hill VR . Am J Trop Med Hyg 2014 92 (1) 118-25 ![]() Epidemic cholera was reported in Haiti in 2010, with no information available on the occurrence or geographic distribution of toxigenic Vibrio cholerae in Haitian waters. In a series of field visits conducted in Haiti between 2011 and 2013, water and plankton samples were collected at 19 sites. Vibrio cholerae was detected using culture, polymerase chain reaction, and direct viable count methods (DFA-DVC). Cholera toxin genes were detected by polymerase chain reaction in broth enrichments of samples collected in all visits except March 2012. Toxigenic V. cholerae was isolated from river water in 2011 and 2013. Whole genome sequencing revealed that these isolates were a match to the outbreak strain. The DFA-DVC tests were positive for V. cholerae O1 in plankton samples collected from multiple sites. Results of this survey show that toxigenic V. cholerae could be recovered from surface waters in Haiti more than 2 years after the onset of the epidemic. |
Improved method for the detection and quantification of Naegleria fowleri in water and sediment using immunomagnetic separation and real-time PCR
Mull BJ , Narayanan J , Hill VR . J Parasitol Res 2013 2013 608367 Primary amebic meningoencephalitis (PAM) is a rare and typically fatal infection caused by the thermophilic free-living ameba, Naegleria fowleri. In 2010, the first confirmed case of PAM acquired in Minnesota highlighted the need for improved detection and quantification methods in order to study the changing ecology of N. fowleri and to evaluate potential risk factors for increased exposure. An immunomagnetic separation (IMS) procedure and real-time PCR TaqMan assay were developed to recover and quantify N. fowleri in water and sediment samples. When one liter of lake water was seeded with N. fowleri strain CDC:V212, the method had an average recovery of 46% and detection limit of 14 amebas per liter of water. The method was then applied to sediment and water samples with unknown N. fowleri concentrations, resulting in positive direct detections by real-time PCR in 3 out of 16 samples and confirmation of N. fowleri culture in 6 of 16 samples. This study has resulted in a new method for detection and quantification of N. fowleri in water and sediment that should be a useful tool to facilitate studies of the physical, chemical, and biological factors associated with the presence and dynamics of N. fowleri in environmental systems. |
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