Guide for Detection of Nucleic Acid Sequences of the Human Immunodeficiency Virus HIV-1 by the Polymerase Chain Reaction Technique

10.1520/e2146 ◽  
2010 ◽  
Author(s):  
Keyword(s):  
Human Immunodeficiency Virus ◽  
Polymerase Chain Reaction ◽  
Nucleic Acid ◽  
Polymerase Chain Reaction Technique ◽  
Chain Reaction ◽  
Immunodeficiency Virus ◽  
Polymerase Chain ◽  
Hiv 1 ◽  
Nucleic Acid Sequences

Analysis of sewage effluent for human immunodeficiency virus (HIV) using infectivity assay and reverse transcriptase polymerase chain reaction

1995 ◽  
Vol 41 (9) ◽  
pp. 809-815 ◽  
Author(s):  
Carol J. Palmer ◽  
G. Fred Bonilla ◽  
Yu-Li Tsai ◽  
Moon H. Lee ◽  
Brenda J. Javier ◽  
...  
Keyword(s):  
Waste Water ◽  
Human Immunodeficiency Virus ◽  
Polymerase Chain Reaction ◽  
Sewage Effluent ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Treated Sewage ◽  
Immunodeficiency Virus ◽  
Polymerase Chain ◽  
Hiv 1

Environmental survival of human immunodeficiency virus type 1 (HIV-1) is an important public health concern. Survival of HIV in waste water is of particular interest to those who work at treatment facilities and to the general public who have contact with rivers or ocean water receiving treated sewage effluent. Other researchers have reported that HIV can be detected in waste water. Their studies, however, detected homologous nucleic acid sequences but did not attempt to determine infectivity. The current study tested primary and secondary effluent from a major metropolitan sewage agency for the presence of HIV-1 using reverse transcriptase polymerase chain reaction (RT-PCR), HIV-1 p24 antigen enzyme-linked immunosorbent assay, and infectivity testing. For RT-PCR, primers SK38/SK39 and M667/AA55 were used to identify HIV-1 RNA sequences from concentrated and extracted sewage samples. Infectivity assays employed donor peripheral blood mononuclear cells (PBMCs) stimulated with phytohemagglutinin. Coxsackievirus B4, echovirus 7, and poliovirus 1, enteroviruses normally present in sewage, were tested for replication in PBMCs. Poliovirus 1 was found to infect the PBMCs. To eliminate other enteroviruses that may also infect the PBMCs and interfere with HIV-1 testing, concentrated sewage was treated with human immunoglobulin (free of HIV antibodies) and poliovirus antisera before infectivity assays were performed. All treated sewage samples tested negative for HIV-1 by all methods used. HIV-1 seeded into sewage, however, remained infectious in the assay, indicating that the sewage water sample did not interfere with HIV infectivity nor was it toxic to the PBMCs.Key words: HIV, sewage, RT-PCR, infectivity.

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