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A multiplex real‐time PCR for quantification of HIV‐1 DNA and the human albumin gene in CD4 + cells
Author(s) -
ERIKSSON LARS E.,
LEITNER THOMAS,
WAHREN BRITTA,
BOSTRÖM ANNCHARLOTTE,
FALK KERSTIN I.
Publication year - 2003
Publication title -
apmis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.909
H-Index - 88
eISSN - 1600-0463
pISSN - 0903-4641
DOI - 10.1034/j.1600-0463.2003.1110605.x
Subject(s) - multiplex , multiplex polymerase chain reaction , dna , biology , microbiology and biotechnology , gene , virology , real time polymerase chain reaction , in vitro , virus , genome , polymerase chain reaction , in vivo , genetics
We have established a simple system for measuring HIV‐1 DNA load in CD4 + cells. In a multiplex configuration, a conserved region in the HIV‐1 pol gene and a section of the human albumin gene were simultaneously amplified to estimate the number of HIV‐1 DNA copies per cellular genome. An established Epstein‐Barr virus (EBV) standard system was used to calibrate the HIV‐1 quantification. Our multiplex PCR system was tested on different in vitro developed HIV‐1 strains and on longitudinal samples from eight patients. The system was able to amplify both in vitro and in vivo samples of various genetic compositions. In all eight patients, HIV‐1 DNA was detected and ranged between 0.17 and 51×10 −3 copies per CD4 + cell and could be monitored longitudinally, including long‐term PI‐ART and STI. The measured HIV‐1 DNA load may be used to select the best time for the institution or re‐institution of therapy.