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废水中与人类免疫缺陷病毒同源的核酸检测。

Detection of nucleic acids homologous to human immunodeficiency virus in wastewater.

作者信息

Preston D R, Farrah S R, Bitton G, Chaudhry G R

机构信息

Department of Microbiology, University of Florida, Gainesville.

出版信息

J Virol Methods. 1991 Aug;33(3):383-90. doi: 10.1016/0166-0934(91)90038-2.

Abstract

Raw wastewaters were obtained from the cities of Belle Glade, Ocala and Gainesville in the state of Florida and were concentrated using several established methods for the recovery of human enteroviruses. The nucleic acids were then extracted from the wastewater concentrates, suspended in 2 x SSC with and without 2 N NaOH (for the detection of DNA and both DNA and RNA, respectively), and dot blotted onto hybridization membranes. These membranes were then hybridized with three 32P-end-labeled 18-mer oligonucleotides directed against the LTR, gag, and env regions of the human immunodeficiency virus type 1 (HIV-1). Autoradiographic analyses of these blots indicate that sequences homologous to HIV-1 genomic RNA and proviral DNA were found in Belle Glade wastewater but not in wastewater from Ocala and Gainesville. These findings may have implications in the wastewater treatment system as well as for detection of HIV-1 in clinical samples.

摘要

原废水取自佛罗里达州的贝尔格莱德市、奥卡拉市和盖恩斯维尔市,采用几种成熟的方法对其进行浓缩,以回收人肠道病毒。然后从浓缩的废水中提取核酸,分别悬浮于含有和不含有2N氢氧化钠的2×SSC中(分别用于检测DNA以及DNA和RNA),并点样于杂交膜上。然后将这些膜与三种针对1型人类免疫缺陷病毒(HIV-1)的长末端重复序列(LTR)、群抗原(gag)和包膜蛋白(env)区域的32P末端标记的18聚体寡核苷酸进行杂交。对这些印迹的放射自显影分析表明,在贝尔格莱德市的废水中发现了与HIV-1基因组RNA和前病毒DNA同源的序列,但在奥卡拉市和盖恩斯维尔市的废水中未发现。这些发现可能对废水处理系统以及临床样本中HIV-1的检测具有重要意义。

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