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The Self‐sustained sequence replication amplification system–move over, PCR
Author(s) -
Lown Kenneth S.
Publication year - 1993
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.1840170229
Subject(s) - biology , virology , peripheral blood mononuclear cell , microbiology and biotechnology , zidovudine , polymerase chain reaction , genotype , nucleic acid sequence , virus , gene , amino acid , peptide sequence , genetics , viral disease , in vitro
Mutations at amino acid positions 67, 70, 215, and 219 in the human immunodeficiency virus type 1 (HIV‐1) pol gene correlate with the emergence of resistance to zidovudine (AZT). These four positions were monitored in viral RNA extracted from infected peripheral blood mononuclear cells (PBMC) and viral stocks obtained after coculture with uninfected lymphocytes. Genotype determinations were made using the selfsustained sequence replication (3SR) and differential bead‐based sandwich hybridization (BBSH) assay. The hybridization results obtained by 3SR and BBSH analyses were verified by dideoxynucleotide sequencing of the 3SR products. Correlation of 3SR and BBSH with the polymerase chain reaction and Southern hybridization analyses of the PBMC and corresponding viral isolates indicated that PBMC and corresponding HIV‐1 isolates may differ in their genotypes at the monitored amino acid positions, variations from the wild‐type nucleotide sequence may occur proximal to the codons being monitored, and viral isolates possessing the same genotypes at the four monitored amino acid positions showed a threefold variation in their ID 50 measurements.