Electronic transduction of HIV-1 drug resistance in AIDS patients.
A drug composition consisting of nucleoside reverse transcriptase inhibitors (NRTIs), non-nucleoside reverse transcriptase inhibitors (NNRTIs), and protease inhibitors (PIs) is commonly used in AIDS therapy. A major difficulty encountered with the therapeutic composite involves the emergence of drug-resistant viruses, especially to the PIs, regarded as the most effective drugs in the composition. We present a novel bioelectronic means to detect the appearance of mutated HIV-1 exhibiting drug resistance to the PI saquinavir. The method is based on the translation of viral RNA, the association of cleaved or uncleaved Gag polyproteins at an electrode surface functionalized with the respective antibodies, and the bioelectronic detection of the Gag polyproteins associated with the surface. The bioelectronic process includes the association of anti-MA or anti-CA antibodies, the secondary binding of an antibody-horseradish peroxidase (HRP) conjugate, and the biocatalyzed precipitation of an insoluble product on the electronic transducers. Faradaic impedance measurements and quartz crystal microbalance analyses are employed to follow the autoprocessing of the Gag polyproteins. The method was applied to determine drug resistance in infected cultured cells and also in blood samples of consenting AIDS patients. The method described here is also applicable to the determination of drug effectiveness in AIDS patients and to screening of the efficiency of newly developed drugs.
Alfonta
Lital
L
Institute of Chemistry, The Hebrew University of Jerusalem, Jerusalem 91904, Israel.
Blumenzweig
Immanuel
I
Zayats
Maya
M
Baraz
Lea
L
Kotler
Moshe
M
Willner
Itamar
I
D019380:Anti-HIV Agents / Q000627:therapeutic use*; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000378:metabolism; D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods*; D002478:Cells, Cultured; D024882:Drug Resistance, Viral / Q000235:genetics*; D004563:Electrochemistry / Q000379:methods; D004566:Electrodes; D004800:Enzymes, Immobilized / Q000737:chemistry*; D015683:Gene Products, gag / Q000737:chemistry / Q000378:metabolism; D005838:Genotype; D015658:HIV Infections / Q000188:drug therapy* / Q000821:virology; D017320:HIV Protease Inhibitors / Q000009:adverse effects; D015497:HIV-1 / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D006801:Humans; D014176:Protein Biosynthesis; D012367:RNA, Viral / Q000032:analysis*; D019258:Saquinavir / Q000009:adverse effects
D019380:Anti-HIV Agents; D000911:Antibodies, Monoclonal; D004800:Enzymes, Immobilized; D015683:Gene Products, gag; D017320:HIV Protease Inhibitors; D012367:RNA, Viral; D019258:Saquinavir