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Development of An Astrovirus Rt-Pcr Detection Assay for Use With Conventional, Real-Time, and Integrated Cell Culture/Rt-Pcr

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Abstract:Astrovirus is a common cause of gastroenteritis in humans that has been determined to be responsible for outbreaks of illness in several countries. Since astrovirus can be waterborne, it is important to be able to identify this virus in environmental water. We have developed and optimized an RT-PCR method that was able to amplify all eight astrovirus serotypes. In addition, an internal control was designed so that any inhibitors of this astrovirus assay could be detected. The assay was adapted for use in a real-time PCR assay and the sensitivity of these two methods was compared. The real-time assay was then combined with CaCo2 cell culture to produce an integrated cell culture/RT-PCR (ICC/RT-PCR) assay and the sensitivity of the combined assay was compared to RT-PCR alone. The methods were used to detect astrovirus in acute phase illness stool samples as well as to detect the virus in water samples spiked with astrovirus.
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Citation:Grimm, A. C., J. L. Cashdollar, F. P. Williams, and G. S. Fout. Development of An Astrovirus Rt-Pcr Detection Assay for Use With Conventional, Real-Time, and Integrated Cell Culture/Rt-Pcr. CANADIAN JOURNAL OF MICROBIOLOGY 50(4):269-278, (2004).
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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: Journal
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Published: 04/01/2004
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Downloads:
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Bullet Item Development of An Astrovirus Rt-Pcr Detection Assay for Use With Conventional, Real-Time, and Integrated Cell Culture/Rt-Pcr
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Related Entries:
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Bullet Item Detecting Ccl-Related, Emerging Waterborne Human Viruses and Viral Indicators for Exposure Assessment
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Last Updated on Monday, October 22, 2007
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