The application of a poliovirus cDNA probe for the detection of enteroviruses in water

A. B. Margolin; M. J. Hewlett; C. P. Gerba

doi:10.2166/wst.1991.0073

The application of a poliovirus cDNA probe for the detection of enteroviruses in water

A. B. Margolin, M. J. Hewlett, C. P. Gerba

Research output: Chapter in Book/Report/Conference proceeding › Conference contribution

4 Scopus citations

Abstract

A technique for the detection of enteroviruses in water using gene probes in a dot blot assay is described. Poliovirus cDNA probes were labeled with [³²P] dCTP and [³²P] dATP to a specific activity greater than 2.0 × 10⁹ cpm/μg DNA. Autoradiograph times were compared to determine the minimum time required to optimize the sensitivity of the test. One infectious unit of poliovirus was detected from cell harvests within 48 hours using gene probes. Potential false positive reactions between bacterial DNA in environmental samples and vector DNA were controlled by both base hydrolysis of viral RNA and enzymatic treatment of the sample. Upon comparison to cell culture, the dot blot assay was as sensitive for the detection of virus in sewage contaminated groundwater.

Original language	English (US)
Title of host publication	Health-Related Water Microbiolgy 1990
Pages	277-280
Number of pages	4
Volume	24
Edition	2
DOIs	https://doi.org/10.2166/wst.1991.0073
State	Published - 1991
Externally published	Yes
Event	Proceedings of the IAWPRC International Symposium - Tuebingen, Ger Duration: Apr 1 1990 → Apr 6 1990

Other

Other	Proceedings of the IAWPRC International Symposium
City	Tuebingen, Ger
Period	4/1/90 → 4/6/90

ASJC Scopus subject areas

Environmental Engineering
Water Science and Technology

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10.2166/wst.1991.0073

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title = "The application of a poliovirus cDNA probe for the detection of enteroviruses in water",

abstract = "A technique for the detection of enteroviruses in water using gene probes in a dot blot assay is described. Poliovirus cDNA probes were labeled with [32P] dCTP and [32P] dATP to a specific activity greater than 2.0 × 109 cpm/μg DNA. Autoradiograph times were compared to determine the minimum time required to optimize the sensitivity of the test. One infectious unit of poliovirus was detected from cell harvests within 48 hours using gene probes. Potential false positive reactions between bacterial DNA in environmental samples and vector DNA were controlled by both base hydrolysis of viral RNA and enzymatic treatment of the sample. Upon comparison to cell culture, the dot blot assay was as sensitive for the detection of virus in sewage contaminated groundwater.",

author = "Margolin, {A. B.} and Hewlett, {M. J.} and Gerba, {C. P.}",

year = "1991",

doi = "10.2166/wst.1991.0073",

language = "English (US)",

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volume = "24",

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T1 - The application of a poliovirus cDNA probe for the detection of enteroviruses in water

AU - Margolin, A. B.

AU - Hewlett, M. J.

AU - Gerba, C. P.

PY - 1991

Y1 - 1991

N2 - A technique for the detection of enteroviruses in water using gene probes in a dot blot assay is described. Poliovirus cDNA probes were labeled with [32P] dCTP and [32P] dATP to a specific activity greater than 2.0 × 109 cpm/μg DNA. Autoradiograph times were compared to determine the minimum time required to optimize the sensitivity of the test. One infectious unit of poliovirus was detected from cell harvests within 48 hours using gene probes. Potential false positive reactions between bacterial DNA in environmental samples and vector DNA were controlled by both base hydrolysis of viral RNA and enzymatic treatment of the sample. Upon comparison to cell culture, the dot blot assay was as sensitive for the detection of virus in sewage contaminated groundwater.

AB - A technique for the detection of enteroviruses in water using gene probes in a dot blot assay is described. Poliovirus cDNA probes were labeled with [32P] dCTP and [32P] dATP to a specific activity greater than 2.0 × 109 cpm/μg DNA. Autoradiograph times were compared to determine the minimum time required to optimize the sensitivity of the test. One infectious unit of poliovirus was detected from cell harvests within 48 hours using gene probes. Potential false positive reactions between bacterial DNA in environmental samples and vector DNA were controlled by both base hydrolysis of viral RNA and enzymatic treatment of the sample. Upon comparison to cell culture, the dot blot assay was as sensitive for the detection of virus in sewage contaminated groundwater.

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U2 - 10.2166/wst.1991.0073

DO - 10.2166/wst.1991.0073

M3 - Conference contribution

AN - SCOPUS:0025761869

SN - 0080411495

SN - 9780080411491

VL - 24

SP - 277

EP - 280

BT - Health-Related Water Microbiolgy 1990

T2 - Proceedings of the IAWPRC International Symposium

Y2 - 1 April 1990 through 6 April 1990

ER -

The application of a poliovirus cDNA probe for the detection of enteroviruses in water

Abstract

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