| 摘要 |
Immunocapture of plasma HIV-1, coupled with direct lysis of the virions and a simplified method of reverse transcription and amplification of the HIV-1 cDNA by the Polymerase Chain Reaction (PCR) represents a rapid and highly sensitive method to monitor HIV-1 disease progression. This method is also less time and labor intensive than quantitative culture. In addition, the development of a method to directly lyse the immunocaptured virions and a simplified single step reverse transcription (RT)/PCR procedure eliminated the need for organic solvent extraction and reduced the number of steps in the procedure. A direct lysis buffer was formulated to isolate plasma HIV-1 RNA for direct use in the RT and PCR reactions, thus eliminating the need for organic solvent extraction and ethanol precipitation. This resulted in a significant saving of time needed to complete the assay and significantly reduces the possibility of contamination associated with PCR reactions. The immunocapture-RT/PCR assay was used to show that vertical transmission of HIV-1 from a mother to her child depended largely on factors other that viral load. Conversely, the plasma viral load played a significant role in transfusion associated transmission of HIV-1 infection. Finally, the detection and quantitation of plasma associated viral load by immunocapture-RT/PCR may provide an additional marker of disease progression and may aid in determining the efficacy of various HIV therapeutics. |
