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废水中人类免疫缺陷病毒核酸的存在及其通过聚合酶链反应进行的检测。

Presence of human immunodeficiency virus nucleic acids in wastewater and their detection by polymerase chain reaction.

作者信息

Ansari S A, Farrah S R, Chaudhry G R

机构信息

Department of Biological Sciences, Oakland University, Rochester, Michigan 48309.

出版信息

Appl Environ Microbiol. 1992 Dec;58(12):3984-90. doi: 10.1128/aem.58.12.3984-3990.1992.

Abstract

The human immunodeficiency virus type 1 (HIV-1) released by infected individuals or present in human and hospital wastes can potentially cause contamination problems. The presence of HIV-1 was investigated in 16 environmental samples, including raw wastewater, sludge, final effluent, soil, and pond water, collected from different locations. A method was developed to extract total nucleic acids in intact form directly from the raw samples or from the viral concentrates of the raw samples. The isolated nucleic acids were analyzed for the presence of HIV-1 by using in vitro amplification of the target sequences by the polymerase chain reaction (PCR) method. HIV-1-specific proviral DNA and viral RNA were detected in the extracted nucleic acids obtained from three wastewater samples by this method. The specificity of the PCR-amplified products was determined by Southern blot hybridization with an HIV-1-specific oligonucleotide probe, SK19. The isolated nucleic acids from wastewater samples were also screened for the presence of poliovirus type 1, representing a commonly found enteric virus, and simian immunodeficiency virus, representing, presumably, rare viruses. While poliovirus type 1 viral RNA was found in all of the wastewater samples, none of the samples yielded a simian immunodeficiency virus-specific product. No PCR-amplified product was yielded when wastewater samples were directly used for the detection of HIV-1 and poliovirus type 1. The wastewater constituents appeared to be inhibitory to the enzymes reverse transcriptase and DNA polymerase.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

受感染个体释放的或存在于人类及医院废弃物中的1型人类免疫缺陷病毒(HIV-1)可能会造成污染问题。对从不同地点采集的16份环境样本进行了HIV-1检测,这些样本包括原废水、污泥、最终出水、土壤和池塘水。开发了一种方法,可直接从原始样本或原始样本的病毒浓缩物中完整提取总核酸。通过聚合酶链反应(PCR)方法对靶序列进行体外扩增,分析分离出的核酸中是否存在HIV-1。用该方法在从3份废水样本中提取的核酸中检测到了HIV-1特异性前病毒DNA和病毒RNA。通过与HIV-1特异性寡核苷酸探针SK19进行Southern印迹杂交,确定了PCR扩增产物的特异性。还对废水样本中分离出的核酸进行了检测,以查找1型脊髓灰质炎病毒(一种常见的肠道病毒)和猴免疫缺陷病毒(大概是罕见病毒)。虽然在所有废水样本中都发现了1型脊髓灰质炎病毒的病毒RNA,但没有一个样本产生猴免疫缺陷病毒特异性产物。直接使用废水样本检测HIV-1和1型脊髓灰质炎病毒时,未产生PCR扩增产物。废水成分似乎对逆转录酶和DNA聚合酶有抑制作用。(摘要截短于250词)

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3187/183215/4cc5eb082c34/aem00053-0212-a.jpg

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