Recombinant envelope protein of HIV-1 subtype E as antigen in HIV-1 antibody detection enzyme immunoassay.

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dc.contributor.authorSutthent, Ruengpungen_US
dc.contributor.authorKanoksinsombat, Chindaen_US
dc.contributor.authorHorthongkham, Navinen_US
dc.contributor.authorLouisirirotchanakul, Sudaen_US
dc.contributor.authorAuewarakul, Praserten_US
dc.contributor.authorKantakamalakul, Wanneeen_US
dc.date.accessioned2009-05-27T16:28:27Z
dc.date.available2009-05-27T16:28:27Z
dc.date.issued2002-06-19en_US
dc.descriptionThe Southeast Asian Journal of Tropical Medicine and Public Health.en_US
dc.description.abstractIn order to develop a reliable and inexpensive serodiagnostic method to be used for anti-HIV antibody detection in Thailand, recombinant envelope (TM or gp41 subunit) protein of HIV-1 subtype E was produced from prokaryotic cell (Escherichia coli) as the source of antigen in enzyme immunoassay (TE diagnostic EIA kit). HIV-1 gp41 subunit of subtype E was successfully expressed in E. coli in the form of polyhistidine-tagged proteins, comprising of rgp41A (601 bases N-terminal half of TM or 25kDa) and rgp41B (560 bases C-terminal half of TM or 24 kDa) by using an expression vector, pBAD/His C. The amount of protein, dilution of sera, and anti-human IgG labeled HRP used in the EIA test optimized by a checker board titration of the protein and seropositive or seronegative sera, were 5.0 microg/ml, 1:300, and 1:4,000, respectively. The blinded test evaluation of TE-diagnostic EIA in 500 seropositive and 500 seronegative sera which have been simultaneously tested by two available commercial kits and compared with our TE diagnostic EIA, gave 99.6% sensitivity and specificity. The other known genetic subtypes sera such as subtype A (n=5), B (n=9), C (n=4) and D (n=5) were also positive with this EIA. The estimated manufacturer cost per test of rgp41 based anti-HIV antibody detection EIA or TE-diagnostic EIA was about 15 baht. This recombinant envelope (gp41 or TM) protein from HIV-1, which can be produced in large quantities without any hazards from growing the virus and has lower cost to produce anti-HIV antibody serological diagnostic kit, should be considered as an HIV screening test in Thailand.en_US
dc.description.affiliationDepartment of Microbiology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand. sirst@mahidol.ac.then_US
dc.identifier.citationSutthent R, Kanoksinsombat C, Horthongkham N, Louisirirotchanakul S, Auewarakul P, Kantakamalakul W. Recombinant envelope protein of HIV-1 subtype E as antigen in HIV-1 antibody detection enzyme immunoassay. The Southeast Asian Journal of Tropical Medicine and Public Health. 2002 Jun; 33(2): 288-96en_US
dc.identifier.urihttps://imsear.searo.who.int/handle/123456789/35290
dc.language.isoengen_US
dc.source.urihttps://www.tm.mahidol.ac.th/seameo/2002_33_2/15-2834.pdfen_US
dc.subject.meshBase Sequenceen_US
dc.subject.meshDNA Primersen_US
dc.subject.meshHIV Antibodies --blooden_US
dc.subject.meshHIV Envelope Protein gp41 --immunologyen_US
dc.subject.meshHIV-1 --classificationen_US
dc.subject.meshHumansen_US
dc.subject.meshImmunoenzyme Techniques --methodsen_US
dc.subject.meshRecombinant Proteins --immunologyen_US
dc.titleRecombinant envelope protein of HIV-1 subtype E as antigen in HIV-1 antibody detection enzyme immunoassay.en_US
dc.typeJournal Articleen_US
dc.typeResearch Support, Non-U.S. Gov'ten_US
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