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LABORATORY PROCEDURES FOR ANIMAL & HUMAN CELL LINES

REFERENCE NO: AHC/1998/3/3.4/1.3


TITLE: PCR ANALYSIS FOR HBV, HCV, HHV-8


The active form of the human pathogenic virus HBV is tested by extracting DNA from the supernatant of the cell lines after enrichment of particles by ultracentrifugation. HCV RNA is prepared from cells by organic extraction followed by DNase digestion. PCR or RT-PCR for HBV and HCV, respectively, is carried out with the same parameters as described for EBV and primers coding for conserved sequences of the X gene of HBV (F-HBV and R-HBV) and from the 5-UTR of HCV (1). A selected number of cell lines has also been tested by PCR for the occurrence of the recently described human herpesvirus type 8 (HHV-8) (2).

References:

1. Pontisso P, Ruvoletto MG, Fattovich G, Chemello L, Gallorini A, Ruol A, Alberti A: Clinical and virological profiles in patients with multiple hepatitis virus infection. Gastroenterology 105: 1529-1533 (1993).

2. Uphoff CC, Habig S, Carbone A, Gaidano G, Drexler HG: HHV-8 infection is specific for cell lines derived from primary effusion (body cavity-based) lymphomas. Leukemia (in press, 1998).


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