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Development of a rapid virus particl...

Ferris, Matthew Marsh.

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Development of a rapid virus particle counter.

紀錄類型:	書目-語言資料,印刷品 : Monograph/item
正題名/作者:	Development of a rapid virus particle counter./
作者:	Ferris, Matthew Marsh.
面頁冊數:	170 p.
附註:	Director: Kathy L. Rowlen.
Contained By:	Dissertation Abstracts International63-02B.
標題:	Biology, Microbiology. -
電子資源:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=3043523
ISBN:	0493573356

Development of a rapid virus particle counter.
Ferris, Matthew Marsh.

Development of a rapid virus particle counter. - 170 p.

Director: Kathy L. Rowlen.

Thesis (Ph.D.)--University of Colorado at Boulder, 2002.

Nanometric biological particles such as viruses have received increased attention in a wide range of scientific fields. Evaluation of viral contributions to environmental processes and the use of viruses in medical applications such as gene therapy, require viruses to be routinely and accurately enumerated (i.e. counted). There are a variety of existing techniques for counting viruses, namely plaque assays, transmission electron microscopy (TEM), epifluorescence microscopy (EFM) and flow cytomtery (FCM); each has advantages and disadvantages. However, the disadvantages associated with current techniques warrant the development of faster methods with increased reliability and sensitivity. The Single Nanometric Particle Enumerator (SNaPE) was developed specifically for counting viruses solution.

ISBN: 0493573356Subjects--Topical Terms:

1017734
Biology, Microbiology.

Development of a rapid virus particle counter.
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245 1 0 $a Development of a rapid virus particle counter.
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500 $a Director: Kathy L. Rowlen.
500 $a Source: Dissertation Abstracts International, Volume: 63-02, Section: B, page: 0773.
502 $a Thesis (Ph.D.)--University of Colorado at Boulder, 2002.
520 $a Nanometric biological particles such as viruses have received increased attention in a wide range of scientific fields. Evaluation of viral contributions to environmental processes and the use of viruses in medical applications such as gene therapy, require viruses to be routinely and accurately enumerated (i.e. counted). There are a variety of existing techniques for counting viruses, namely plaque assays, transmission electron microscopy (TEM), epifluorescence microscopy (EFM) and flow cytomtery (FCM); each has advantages and disadvantages. However, the disadvantages associated with current techniques warrant the development of faster methods with increased reliability and sensitivity. The Single Nanometric Particle Enumerator (SNaPE) was developed specifically for counting viruses solution.
520 $a The SNaPE instrument utilizes capillary-based flow cytometry and confocal single molecule detection optics to achieve rapid, accurate and reliable detection of individual fluorescent particles. The SNaPE was characterized using fluorescent polystryrene nanospheres with diameters and fluorescence intensities similar to those expected from a virus stained with a nucleic acid dye. A systematic investigation of data acquisition parameters was conducted to ensure accurate results under optimized conditions. A quantitative comparison of SNaPE, TEM, EFM and FCM showed SNaPE and EFM to have the highest degree of accuracy and precision for particle concentrations between 10<super>6</super> and 10<super> 9</super> per mL. Additionally, the fluorescence limit of detection for the SNaPE instrument was experimentally determined to be 180 FITC equivalents; a value &sim;2.5x lower than that of the commercial flow cytometer examined in the comparison.
520 $a The reliability of the SNaPE instrument with respect to virus enumeration was demonstrated for three medically relevant viruses, adenovirus-5, respiratory syncytial virus and influenza A, treated with a fluorescent nucleotide stain. In each case, the SNaPE yielded virus particle concentrations consistent with, but slightly lower than TEM results, as expected. In addition, based on calibration of signal intensity, the average peak height for a given virus was highly correlated with genome size. In contrast to time consuming analyses by TEM and plaque titers, SNaPE analysis of pure virus samples (including sample handling, data collection and data processing) can be completed within one hour. The SNaPE represents a rapid and inexpensive alternative method for virus enumeration.
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791 $a Ph.D.
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