A rapid method for comparative quantitative polymerase chain reaction of HIV-1 proviral DNA extracted from cryopreserved brain tissues.
The quantitative polymerase chain reaction (PCR) method devised by Fujimura and Bockstahler (1995) was modified for rapid determination of distribution of HIV-1 proviral DNA load in AIDS brains. It was used for analysis of an association with HIV-1 associated dementia and HIV-1 encephalitis (Fujimura et al., 1997). The method has wider applicability for comparative studies of viral DNA load based on PCR amplification. The method is applicable under conditions where target DNA and its PCR-amplified product increase proportionally. An equation was derived to obtain the number of copies of HIV-1 DNA per cellular genome from the amount of PCR amplified product of a tissue specimen DNA. The equalizing constant is the reciprocal of the slope of the amplification of the HIV-1 proviral DNA sequence of the standard cellular DNA included in each experiment. The intercept of the equation is zero.
Fujimura
R K
RK
Department of Psychiatry, University of Miami School of Medicine, FL, USA.
Shapshak
P
P
Feaster
D
D
Epler
M
M
Goodkin
K
K
D001921:Brain / Q000473:pathology / Q000821:virology*; D015925:Cryopreservation; D004279:DNA, Viral / Q000302:isolation & purification*; D018628:Gene Dosage; D016679:Genome, Viral; D015658:HIV Infections / Q000821:virology; D015497:HIV-1 / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D011533:Proviruses / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D015203:Reproducibility of Results; D019562:Viral Load
D004279:DNA, Viral