Strategies for detection of transfusion-transmitted viruses eluding identification by conventional serologic tests. II. Detection of host DNA in human plasmas with elevated alanine aminotransferase.
As a prelude to the development of nucleic acid probes specific for non-A, non-B hepatitis virus(es) (NANBV), plasmas with alanine aminotransferase levels greater than or equal to 110 IU were assayed for DNA by a radioimmunoassay. Approximately 50% of such plasmas are expected to contain NANBV. One-hundred and seventy-eight of 420 plasma samples tested (42.4%) contained sequestered DNA resistant to DNAse I. The DNA has a molecular weight of approximately equal to 0.8 to 1.4 X 10(6) daltons and hybridizes with a 32P-labeled human DNA probe. The DNA in plasma is mostly bound to IgM. The presence of host DNA will have to be taken into account in planning experiments aiming at the preparation of nucleic acid probes specific for NANBV using infected plasmas as source material. Such experiments will have to utilize recombinant DNA technology and will require the separation of bacterial colonies containing recombinant DNA with viral DNA sequences from colonies with human DNA inserts. The feasibility of this approach is demonstrated using plasma-containing hepatitis B virus.
D000410:Alanine Transaminase / Q000097:blood*; D002499:Centrifugation, Density Gradient; D003001:Cloning, Molecular; D004247:DNA / Q000097:blood* / Q000235:genetics / Q000378:metabolism; D004262:DNA Restriction Enzymes; D004274:DNA, Recombinant; D004279:DNA, Viral / Q000097:blood*; D006526:Hepatitis C / Q000382:microbiology; D006518:Hepatitis Viruses / Q000235:genetics*; D006801:Humans; D007075:Immunoglobulin M / Q000378:metabolism; D009693:Nucleic Acid Hybridization; D014176:Protein Biosynthesis; D011863:Radioimmunoassay
D004274:DNA, Recombinant; D004279:DNA, Viral; D007075:Immunoglobulin M; D004247:DNA; D000410:Alanine Transaminase; D004262:DNA Restriction Enzymes