DEVELOPMENT OF PCR METHODS FOR ENTERIC VIRUS DETECTION IN WATER

被引：33

作者：

SCHWAB, KJ

DELEON, R

SOBSEY, MD

机构：

来源：

WATER SCIENCE AND TECHNOLOGY | 1993年 / 27卷 / 3-4期

关键词：

RT-PCR; BEEF EXTRACT; ENTERIC VIRUSES; CONCENTRATION; PURIFICATION; PEG PRECIPITATION; SPIN-COLUMN CHROMATOGRAPHY;

D O I：

10.2166/wst.1993.0348

中图分类号：

X [环境科学、安全科学];

学科分类号：

08 ; 0830 ;

摘要：

This study developed a reverse transcriptase-polymerase chain reaction (RT-PCR) detection system for enteric viruses in sample concentrates obtained by conventional filter adsorption-elution methods. one liter beef extract (BE)-glycine (G) eluant seeded with poliovirus 1 and hepatitis A virus (HAV) was used as a model system and the eluant further processed for RT-PCR compatibility. Sample concentration and purification procedures consisted of polyethylene glycol (PEG) precipitation, Pro-Cipitate (Affinity Technology, New Brunswick, NJ) precipitation, a second PEG precipitation, spin-column chromatography, and ultrafiltration. Sample volumes are reduced from 1 L to 20-40 muL and purified sufficiently for viral detection by RT-PCR. As little as 3 PFU of poliovirus 1 in an initial 1 L eluate were detected by RT-PCR.

引用

页码：211 / 218

页数：8

共 17 条

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