A rapid method for comparative quantitative polymerase chain reaction of HIV-1 proviral DNA extracted from cryopreserved brain tissues

Robert K. Fujimura; Paul Shapshak; Daniel Feaster; Mark Epler; Karl Goodkin

doi:10.1016/S0166-0934(97)00094-3

A rapid method for comparative quantitative polymerase chain reaction of HIV-1 proviral DNA extracted from cryopreserved brain tissues

Robert K. Fujimura, Paul Shapshak, Daniel Feaster, Mark Epler, Karl Goodkin

Research output: Contribution to journal › Article › peer-review

8 Scopus citations

Abstract

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.

Original language	English (US)
Pages (from-to)	177-187
Number of pages	11
Journal	Journal of Virological Methods
Volume	67
Issue number	2
DOIs	https://doi.org/10.1016/S0166-0934(97)00094-3
State	Published - Sep 1997
Externally published	Yes

Keywords

AIDS brains
HIV-1
Proviral DNA
Quantitative polymerase chain reaction (PCR)

ASJC Scopus subject areas

Virology

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10.1016/S0166-0934(97)00094-3

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title = "A rapid method for comparative quantitative polymerase chain reaction of HIV-1 proviral DNA extracted from cryopreserved brain tissues",

abstract = "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.",

keywords = "AIDS brains, HIV-1, Proviral DNA, Quantitative polymerase chain reaction (PCR)",

author = "Fujimura, {Robert K.} and Paul Shapshak and Daniel Feaster and Mark Epler and Karl Goodkin",

note = "Funding Information: We thank the following people for their participation in this study. Brain tissues were obtained by Jocely Gonzalez and Vincent Iglesias from Medical Examiner's Offices in South Florida. Sara Meinz performed the experiments on HIV-1 DNA load. Dr L. Vitkovic, NIMH, Dr L. Bockstahler, FDA, Dr Mauricio Concha, Department of Neurology of this school, and Dr D. Segal, this laboratory, had reviewed the manuscript during its preparation. We thank Dr M.L. King, Department of Cell Biology and Anatomy for the use of her phosphorimager. Research was supported in part by NIDA grants DA04787 and DA07909 to PS. RKF and ME were supported by DA04787S2 from NIDA. ",

year = "1997",

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doi = "10.1016/S0166-0934(97)00094-3",

language = "English (US)",

volume = "67",

pages = "177--187",

journal = "Journal of Virological Methods",

issn = "0166-0934",

publisher = "Elsevier B.V.",

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T1 - A rapid method for comparative quantitative polymerase chain reaction of HIV-1 proviral DNA extracted from cryopreserved brain tissues

AU - Fujimura, Robert K.

AU - Shapshak, Paul

AU - Feaster, Daniel

AU - Epler, Mark

AU - Goodkin, Karl

N1 - Funding Information: We thank the following people for their participation in this study. Brain tissues were obtained by Jocely Gonzalez and Vincent Iglesias from Medical Examiner's Offices in South Florida. Sara Meinz performed the experiments on HIV-1 DNA load. Dr L. Vitkovic, NIMH, Dr L. Bockstahler, FDA, Dr Mauricio Concha, Department of Neurology of this school, and Dr D. Segal, this laboratory, had reviewed the manuscript during its preparation. We thank Dr M.L. King, Department of Cell Biology and Anatomy for the use of her phosphorimager. Research was supported in part by NIDA grants DA04787 and DA07909 to PS. RKF and ME were supported by DA04787S2 from NIDA.

PY - 1997/9

Y1 - 1997/9

N2 - 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.

AB - 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.

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A rapid method for comparative quantitative polymerase chain reaction of HIV-1 proviral DNA extracted from cryopreserved brain tissues

Abstract

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