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DETECTION OF HIV-1 RNA/DNA AND CD4 MRNA IN FECES AND URINE SAMPLES OF THE MULTICENTER AIDS COHORT STUDY VOLUNTEERS

Chakrabarti, Ayan K (2009) DETECTION OF HIV-1 RNA/DNA AND CD4 MRNA IN FECES AND URINE SAMPLES OF THE MULTICENTER AIDS COHORT STUDY VOLUNTEERS. Master's Thesis, University of Pittsburgh. (Unpublished)

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Abstract

HIV infects and depletes CD4+ T cells in Gut Associated Lymphoid Tissues (GALT) of the Gastrointestinal (GI) tract at a very early stage of infection. Furthermore, GALT are the major reservoirs of HIV-1 and may constantly shed virus and CD4+ T cells into the intestinal lumen throughout the entire course of infection. We hypothesize that the dynamic changes of HIV-1 and CD4+ T cell quantities in feces are linked to disease progression and can be used to predict disease prognosis. The aims of this study are to establish sensitive methods for detection and quantitation of HIV-1 and CD4 mRNA in feces, and to use the methods to monitor the amount of HIV-1 RNA/DNA and CD4 mRNA in feces samples of HIV infected patients and to correlate the findings with disease progression. In addition, since urine may potentially serve as a vehicle for HIV-1 transmission we have also measured HIV-1 RNA/DNA in the urine samples from the same population used for the feces study. Our results showed that using normal feces spiked with known copies of DNA and RNA, as low as 2.5 copies of HIV-1 DNA and 40 copies of HIV-1 RNA were detected per input in both nested PCR and RT-nested PCR reactions respectively. Human CD4 mRNA was also detected in feces. From HIV-1 infected volunteers of the Multicenter AIDS Cohort Study (MACS), HIV-1 DNA, RNA and human CD4 mRNA was detected in 8%, 19% and 31%, respectively, in the feces samples from patients with detectable viral load in blood. In the urine samples from the same study population, HIV-1 DNA was detected in 26% of HIV-1 infected donors and this detection is not always correlated with the presence of detectable viral load in blood. This study has major Public health significance as it demonstrates that HIV-1 RNA/DNA could be detected in feces and urine samples, which may lead to the development of a future non-invasive approach to evaluate disease progression and prognosis. In addition, our study demonstrated, for the first time, the presence of human CD4 mRNA in fecal specimens of infected donors, which could be used as a valuable tool in the future to assess the pathogenesis of Gut Associated Lymphoid Tissue over the course of HIV-1 infection.


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Details

Item Type: University of Pittsburgh ETD
Status: Unpublished
Creators/Authors:
CreatorsEmailPitt UsernameORCID
Chakrabarti, Ayan Kayanraju@yahoo.com
ETD Committee:
TitleMemberEmail AddressPitt UsernameORCID
Committee ChairChen, Yuecheny@pitt.eduCHENY
Committee MemberRinaldo, Charles Rrinaldo@pitt.eduRINALDO
Committee MemberGupta, Phalgunipgupta1@pitt.eduPGUPTA1
Committee MemberRiddler, Sharonriddler@msx.dept-med.pitt.eduRIDDLER
Date: 29 June 2009
Date Type: Completion
Defense Date: 27 March 2009
Approval Date: 29 June 2009
Submission Date: 5 April 2009
Access Restriction: 5 year -- Restrict access to University of Pittsburgh for a period of 5 years.
Institution: University of Pittsburgh
Schools and Programs: Graduate School of Public Health > Infectious Diseases and Microbiology
Degree: MS - Master of Science
Thesis Type: Master's Thesis
Refereed: Yes
Uncontrolled Keywords: ; AIDS; CD4+T cell; Feces; HIV-1; PCR; Urine
Other ID: http://etd.library.pitt.edu/ETD/available/etd-04052009-214806/, etd-04052009-214806
Date Deposited: 10 Nov 2011 19:34
Last Modified: 15 Nov 2016 13:38
URI: http://d-scholarship.pitt.edu/id/eprint/6775

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