Improved Method for the Detection and Quantification of Naegleria fowleri in Water and Sediment Using Immunomagnetic Separation and Real-Time PCR
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 th...
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| Format: | Article |
| Language: | English |
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Wiley
2013-01-01
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| Series: | Journal of Parasitology Research |
| Online Access: | http://dx.doi.org/10.1155/2013/608367 |
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| author | Bonnie J. Mull Jothikumar Narayanan Vincent R. Hill |
| author_facet | Bonnie J. Mull Jothikumar Narayanan Vincent R. Hill |
| author_sort | Bonnie J. Mull |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-176233126582441790585e7bf40537f6 |
| institution | Kabale University |
| issn | 2090-0023 2090-0031 |
| language | English |
| publishDate | 2013-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Journal of Parasitology Research |
| spelling | doaj-art-176233126582441790585e7bf40537f62025-02-03T05:45:08ZengWileyJournal of Parasitology Research2090-00232090-00312013-01-01201310.1155/2013/608367608367Improved Method for the Detection and Quantification of Naegleria fowleri in Water and Sediment Using Immunomagnetic Separation and Real-Time PCRBonnie J. Mull0Jothikumar Narayanan1Vincent R. Hill2Division of Foodborne, Waterborne and Environmental Diseases, Centers for Disease Control and Prevention, National Center for Emerging and Zoonotic Infectious Diseases, 1600 Clifton Road, NE, Mail Stop D-66, Atlanta, GA 30329-4018, USADivision of Foodborne, Waterborne and Environmental Diseases, Centers for Disease Control and Prevention, National Center for Emerging and Zoonotic Infectious Diseases, 1600 Clifton Road, NE, Mail Stop D-66, Atlanta, GA 30329-4018, USADivision of Foodborne, Waterborne and Environmental Diseases, Centers for Disease Control and Prevention, National Center for Emerging and Zoonotic Infectious Diseases, 1600 Clifton Road, NE, Mail Stop D-66, Atlanta, GA 30329-4018, USAPrimary 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.http://dx.doi.org/10.1155/2013/608367 |
| spellingShingle | Bonnie J. Mull Jothikumar Narayanan Vincent R. Hill Improved Method for the Detection and Quantification of Naegleria fowleri in Water and Sediment Using Immunomagnetic Separation and Real-Time PCR Journal of Parasitology Research |
| title | Improved Method for the Detection and Quantification of Naegleria fowleri in Water and Sediment Using Immunomagnetic Separation and Real-Time PCR |
| title_full | Improved Method for the Detection and Quantification of Naegleria fowleri in Water and Sediment Using Immunomagnetic Separation and Real-Time PCR |
| title_fullStr | Improved Method for the Detection and Quantification of Naegleria fowleri in Water and Sediment Using Immunomagnetic Separation and Real-Time PCR |
| title_full_unstemmed | Improved Method for the Detection and Quantification of Naegleria fowleri in Water and Sediment Using Immunomagnetic Separation and Real-Time PCR |
| title_short | Improved Method for the Detection and Quantification of Naegleria fowleri in Water and Sediment Using Immunomagnetic Separation and Real-Time PCR |
| title_sort | improved method for the detection and quantification of naegleria fowleri in water and sediment using immunomagnetic separation and real time pcr |
| url | http://dx.doi.org/10.1155/2013/608367 |
| work_keys_str_mv | AT bonniejmull improvedmethodforthedetectionandquantificationofnaegleriafowleriinwaterandsedimentusingimmunomagneticseparationandrealtimepcr AT jothikumarnarayanan improvedmethodforthedetectionandquantificationofnaegleriafowleriinwaterandsedimentusingimmunomagneticseparationandrealtimepcr AT vincentrhill improvedmethodforthedetectionandquantificationofnaegleriafowleriinwaterandsedimentusingimmunomagneticseparationandrealtimepcr |