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Spot‐blot hybridization assay for the detection of hepatitis b virus dna in serum: Factors determining its sensitivity and specificity
Author(s) -
Walter Eike,
Blum Hubert E.,
Offensperger WolfBernhard,
Zeschnigk Christine,
Offensperger Silke,
Gerok Wolfgang
Publication year - 1987
Publication title -
hepatology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.488
H-Index - 361
eISSN - 1527-3350
pISSN - 0270-9139
DOI - 10.1002/hep.1840070323
Subject(s) - dot blot , microbiology and biotechnology , southern blot , hepatitis b virus , virus , virology , hepatitis b virus dna polymerase , biology , dna , chemistry , biochemistry
Factors determining the sensitivity and specificity of the spot‐blot hybridization technique for the detection of hepatitis B virus DNA in serum were systematically investigated. Methods for pretreatment of serum samples, mode of application of the samples to the transfer membranes, blot treatment and hybridization conditions were all found to affect the sensitivity of the assay. The optimum hybridization procedure was found to be incubation of serum samples with salt, NaOH, formaldehyde and detergent, followed by spot application of the samples. This method specifically detected hepatitis B virus DNA in serum with a sensitivity 5 to 15 times higher than the presently used assay procedures.