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Development of a Molecular Method to Identify Astrovirus in Water
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| Abstract: | Astrovirus is a common cause of gastroenteritis that has been determined to be responsible for several outbreaks. Since astrovirus can be waterborne, there is interest in testing environmental water for astrovirus. We have developed a sensitive reverse transcription-polymerase chain reaction (RT-PCR) assay that is designed to detect all known astrovirus strains.
The assay was based on a primer set that contained multiple upper and lower primers as well as multiple probes. This would allow for amplification of all of the known strains of astrovirus using a single reaction. When tested, this assay was able to detect strains from all eight serotypes. In addition, an internal control was developed so that it will be possible to determine if the sample being tested contains PCR inhibitors. Most probable number analysis determined that when amplified with the developed assay, a single DNA molecule of the internal control could be detected if inhibitors were not present. The assay was successfully adapted to real-time PCR and this method was then used for integrated cell culture/RT-PCR detection of infectious virus. The methods were successfully used to detect astrovirus present in clinical samples and spiked water samples.
A simple, sensitive method for detecting all known astrovirus strains has been developed that can be used to detect this virus in water. This assay will be field tested by analyzing environmental water samples. |
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| Citation: | Grimm, A. C., J. L. Cashdollar, F. P. Williams, and G. S. Fout. Development of a Molecular Method to Identify Astrovirus in Water. Presented at USEPA Microorganisms in Drinking Water Workshop, Cincinnati, OH, August 5-7, 2003. |
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| Contact: |
Mary P. O'Bryant - (919)-541-4871 or obriant.mary@epa.gov
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| Division: |
Microbiological & Chemical Exposure Assessment Division |
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| Branch: |
Biohazard Assessment Research Branch |
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| Product Type: |
Abstrct/Oral |
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| Presented: |
08/05/2003 |
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