C - Chemistry – Metallurgy – 12 – Q
Patent
C - Chemistry, Metallurgy
12
Q
C12Q 1/70 (2006.01) C12N 1/06 (2006.01) C12N 7/00 (2006.01) C12N 9/50 (2006.01) C12N 15/10 (2006.01) C12Q 1/68 (2006.01) C12S 3/02 (2006.01)
Patent
CA 2161337
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- I 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.
Henrard Denis R.
Phillips Jack
Abbott Laboratories
Swabey Ogilvy Renault
LandOfFree
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