Irene M Gonzalez

US Patent:

20040072148, Apr 15, 2004

Filed:

Apr 7, 2003

Appl. No.:

10/407897

Inventors:

Jiuping Ji - Rockville MD, US
Mark Manak - Laurel MD, US
Irene Gonzalez - Baltimore MD, US

International Classification:

C12Q001/70
C12P019/34

US Classification:

435/005000, 435/091200

Abstract:

The present invention is directed to a capture assay to simultaneously screen for HBV, HCV and HIV nucleic acids in samples such as plasma. The nucleic acids including both viral DNA and RNA are purified from the plasma samples in a single extraction procedure. In one embodiment, a mixture of degenerate biotin-labelled PCR primers specific for the HBV, HCV, HIV-1 type M and HIV-1 type O are used to amplify any of these viruses which may be present in plasma. Amplified products are captured by hybridization to immobilized capture sequence, and thereafter detected. An internal control vector containing a synthetic fragment flanked by sequences corresponding to the HBV primers was designed to monitor sample recovery during extraction, amplification and detection. All major subtypes of HBV, HCV and HIV including HIV-1 type O have been confirmed and detected by the assay.