Kittigul L, Raengsakulrach B, Siritantikorn S, Kanyok R, Utrarachkij F, Diraphat P, Thirawuth V, Siripanichgon K, Pungchitton S, Chitpirom K, Chaichantanakit N, Vathanophas K
Department of Microbiology, Faculty of Public Health, Mahidol University, Bangkok, Thailand.
Southeast Asian J Trop Med Public Health. 2000 Mar;31(1):41-6.
A modified adsorption-elution technique for concentration of enteric viruses from sewage and water samples was developed. The viruses in water were concentrated by negatively charged membrane filtration, eluted with 2.9% tryptose phosphate broth containing 6% glycine pH 9.0, and reconcentrated using centrifugation by a speedVac concentrator. The presence of poliovirus, hepatitis A virus (HAV) RNA, and rotavirus antigen was determined by cell culture isolation, nested polymerase chain reaction (nested PCR), and enzyme-linked immunosorbent assay (ELISA), respectively. A total of 100 sewage and water samples were collected from various sources in congested communities in Bangkok, concentrated and examined for those enteric viruses. Of 20 surface water samples from canals which located near sewage drains, 15% were positive for HAV RNA by nested PCR. Of 48 domestic sewage samples from man-holes of underground sewers, 8% were positive for rotavirus antigen by ELISA. Even though the samples were concentrated 256-2,000 fold, poliovirus was not found by isolation in cell culture.
开发了一种改进的吸附-洗脱技术,用于从污水和水样中浓缩肠道病毒。水中的病毒通过带负电荷的膜过滤进行浓缩,用含6%甘氨酸、pH 9.0的2.9%胰蛋白胨磷酸盐肉汤洗脱,并用SpeedVac浓缩器通过离心进行再浓缩。分别通过细胞培养分离、巢式聚合酶链反应(巢式PCR)和酶联免疫吸附测定(ELISA)来检测脊髓灰质炎病毒、甲型肝炎病毒(HAV)RNA和轮状病毒抗原的存在。从曼谷拥挤社区的不同来源总共收集了100份污水和水样,对这些水样进行浓缩并检测上述肠道病毒。在来自靠近污水排放口的运河的20份地表水样本中,通过巢式PCR检测发现15%的样本HAV RNA呈阳性。在来自地下污水管道检修孔的48份生活污水样本中,通过ELISA检测发现8%的样本轮状病毒抗原呈阳性。尽管样本被浓缩了256至2000倍,但在细胞培养中未通过分离发现脊髓灰质炎病毒。