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Journal of Clinical Microbiology, April 2000, p. 1414-1418, Vol. 38, No. 4
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Quantitation of Human Immunodeficiency Virus Type 1 RNA in Different Biological Compartments

Robin N. Shepard,1 Jody Schock,1 Kevin Robertson,2 Diane C. Shugars,1,3 John Dyer,4 Pietro Vernazza,5 Colin Hall,2 Myron S. Cohen,6 and Susan A. Fiscus1,*

Department of Microbiology and Immunology,1 Department of Neurology,2 Department of Dental Ecology,3 and Department of Medicine,6 University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599-7140; St. Andrews Hospital, Ipswich, Queensland, Australia4; and Institute for Clinical Microbiology and Immunology, St. Gallen, Switzerland5

Received 12 October 1999/Returned for modification 28 December 1999/Accepted 31 January 2000

Little information is available describing viral loads in body fluids other than blood. In addition, the suitability of commercially available assays for human immunodeficiency virus type 1 (HIV-1) RNA quantitation has not been evaluated in most nonblood fluids. We compared Organon Teknika's nucleic acid sequence-based amplification method (NASBA) and Roche's Amplicor HIV-1 Monitor (reverse transcriptase PCR [RT-PCR]) for quantitating HIV-1 RNA in cerebrospinal fluid (CSF), saliva, breast milk, seminal plasma, and cervical-vaginal lavage fluid (CVL). Saliva and breast milk frequently demonstrated some inhibition in the RT-PCR assay, similar to the inhibition previously described in seminal plasma. Inhibition of the RT-PCR assay was not observed with CSF or CVL, nor in any of the NASBA assays. When fluids from HIV-infected individuals were tested by RT-PCR and NASBA, 73 and 27% of CSF samples and 60 and 40% of breast milk specimens had detectable RNA, respectively. These differences were not statistically significant. In cross-sectional studies using RT-PCR to measure viral RNA in paired blood plasma and CSF samples, 71% of blood plasma samples and 42% of CSF samples were positive. A similar analysis using NASBA with paired blood plasma and CVL, saliva, or seminal plasma samples revealed 91% were blood plasma positive and 55% were CVL positive, 76% were blood plasma positive and 46% were saliva positive, and 83% were blood plasma positive and 63% were seminal plasma positive. NASBA worked fairly well to quantitate HIV-1 RNA from all fluids without apparent inhibition. RT-PCR performed well on CVL and CSF, frequently with greater sensitivity, although its use in other fluids appears limited due to the presence of inhibitors. These studies demonstrate that viral loads in nonblood fluids were generally lower than in blood.

* Corresponding author. Mailing address: University of North Carolina at Chapel Hill Department of Microbiology and Immunology, CB 7140, Chapel Hill, NC 27599-7140. Phone: (919) 966-9872. Fax: (919) 966-9873. E-mail: fiscussa{at}med.unc.edu.

Journal of Clinical Microbiology, April 2000, p. 1414-1418, Vol. 38, No. 4
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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