Jothikumar N, Cliver D O, Mariam T W
Department of Population Health and Reproduction, School of Veterinary Medicine, University of California, Davis 95616-8743, USA.
Appl Environ Microbiol. 1998 Feb;64(2):504-8. doi: 10.1128/AEM.64.2.504-508.1998.
We studied the concentration of hepatitis A virus (HAV) from environmental samples by membrane filter-based urea-arginine phosphate buffer and its detection by using immunomagnetic capture (IC) reverse transcription (RT)-PCR (IC PCR). Magnetic beads coated with anti-HAV rabbit antibodies were used for enrichment and concentration of HAV from environmental samples. IC PCR is sensitive enough to detect as few as 0.04 PFU of cell culture-adapted HAV in inoculated water and sewage samples. IC PCR is specific and does not yield positive reactions with poliovirus 1, HAV RNA, or selected bacteriophages. IC concentrates viruses suspended in small volumes to microliter volumes that can be used directly in RT-PCR. IC concentration of viruses from sewage samples without concentration of inhibitory substances is important for successful RT-PCR detection. In a field trial, 2 of 18 raw sewage samples tested by IC PCR were positive for HAV.
我们通过基于膜滤器的磷酸脲 - 精氨酸缓冲液研究了环境样本中甲型肝炎病毒（HAV）的浓度，并使用免疫磁捕获（IC）逆转录（RT）-PCR（IC PCR）对其进行检测。用抗HAV兔抗体包被的磁珠用于从环境样本中富集和浓缩HAV。IC PCR足够灵敏，能够检测接种水和污水样本中低至0.04 PFU的细胞培养适应型HAV。IC PCR具有特异性，与脊髓灰质炎病毒1、HAV RNA或选定的噬菌体不产生阳性反应。IC将悬浮在小体积中的病毒浓缩至微升体积，可直接用于RT-PCR。在不浓缩抑制性物质的情况下，从污水样本中进行IC病毒浓缩对于成功进行RT-PCR检测很重要。在一项现场试验中，通过IC PCR检测的18份未经处理的污水样本中有2份HAV呈阳性。
