This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Holodniy, M Articles by Merigan, T C Search for Related Content PubMed PubMed Citation Articles by Holodniy, M Articles by Merigan, T C

Inhibition of human immunodeficiency virus gene amplification by heparin.

Center for AIDS Research, Stanford University Medical Center, California 94305.

ABSTRACT

Gene amplification of virus-specific sequences is widely used as a method to detect or confirm human immunodeficiency virus (HIV) infection. In this study we used an enzyme-linked affinity assay to quantify polymerase chain reaction products from whole blood, plasma, and separated mononuclear cells collected in the presence of four common anticoagulants: acid citrate dextrose, sodium EDTA, potassium oxalate, and sodium heparin. Attenuation of the product signal was observed after amplification of nucleic acid extraction from whole blood, washed mononuclear cells, and plasma from specimens collected in sodium heparin. These inhibitory effects on gene amplification could be reversed with heparinase. The addition of as little as 0.05 U of heparin completely inhibited amplification of an HLA-DQa sequence from placental DNA. We conclude that heparin can cause attenuation or inhibition of gene amplification. Acid citrate dextrose and EDTA, which lack inhibitory activity, are the most appropriate anticoagulants for clinical blood samples when polymerase chain reaction amplification is anticipated.

This article has been cited by other articles:

• Sakiyama, R., Fukuta, K., Matsumoto, K., Furukawa, M., Takahashi, Y., Nakamura, T. (2007). Stimulation of Hepatocyte Growth Factor Production by Heparin-derived Oligosaccharides. J Biochem 141: 653-660 [Abstract] [Full Text]  
• Khalesi, B., Bonne, N., Stewart, M., Sharp, M., Raidal, S. (2005). A comparison of haemagglutination, haemagglutination inhibition and PCR for the detection of psittacine beak and feather disease virus infection and a comparison of isolates obtained from loriids. J. Gen. Virol. 86: 3039-3046 [Abstract] [Full Text]  
• Pahwa, R., Hedau, S., Jain, S., Jain, N., Arora, V. M, Kumar, N., Das, B. C (2005). Assessment of possible tuberculous lymphadenopathy by PCR compared to non-molecular methods. J Med Microbiol 54: 873-878 [Abstract] [Full Text]  
• Segura-Alvarez, M., Richter, H., Conraths, F. J., Geue, L. (2003). Evaluation of Enzyme-Linked Immunosorbent Assays and a PCR Test for Detection of Shiga Toxins for Shiga Toxin-Producing Escherichia coli in Cattle Herds. J. Clin. Microbiol. 41: 5760-5763 [Abstract] [Full Text]  
• Kleiboeker, S. B. (2003). Applications of Competitor RNA in Diagnostic Reverse Transcription-PCR. J. Clin. Microbiol. 41: 2055-2061 [Abstract] [Full Text]  
• Kuritzkes, D. R., Grant, R. M., Feorino, P., Griswold, M., Hoover, M., Young, R., Day, S., Lloyd, R. M. Jr., Reid, C., Morgan, G. F., Winslow, D. L. (2003). Performance Characteristics of the TRUGENE HIV-1 Genotyping Kit and the Opengene DNA Sequencing System. J. Clin. Microbiol. 41: 1594-1599 [Abstract] [Full Text]  
• Liu, H., Rodes, B., Chen, C.-Y., Steiner, B. (2001). New Tests for Syphilis: Rational Design of a PCR Method for Detection of Treponema pallidum in Clinical Specimens Using Unique Regions of the DNA Polymerase I Gene. J. Clin. Microbiol. 39: 1941-1946 [Abstract] [Full Text]  
• Kessler, H. H., Stelzl, E., Daghofer, E., Santner, B. I., Marth, E., Lackner, H., Stauber, R. E. (2000). Semiautomated Quantification of Hepatitis B Virus DNA in a Routine Diagnostic Laboratory. CVI 7: 853-855 [Abstract] [Full Text]  
• Kirstein, L. M., Mellors, J. W., Rinaldo, C. R. Jr., Margolick, J. B., Giorgi, J. V., Phair, J. P., Dietz, E., Gupta, P., Sherlock, C. H., Hogg, R., Montaner, J. S. G., Muñoz, A. (1999). Effects of Anticoagulant, Processing Delay, and Assay Method (Branched DNA versus Reverse Transcriptase PCR) on Measurement of Human Immunodeficiency Virus Type 1 RNA Levels in Plasma. J. Clin. Microbiol. 37: 2428-2433 [Abstract] [Full Text]  
• Gilbert, S. A., Burton, K. M., Prins, S. E., Deregt, D. (1999). Typing of Bovine Viral Diarrhea Viruses Directly from Blood of Persistently Infected Cattle by Multiplex PCR. J. Clin. Microbiol. 37: 2020-2023 [Abstract] [Full Text]  
• Tham, J. M., Lee, S. H., Tan, T. M. C., Ting, R. C. Y., Kara, U. A. K. (1999). Detection and Species Determination of Malaria Parasites by PCR: Comparison with Microscopy and with ParaSight-F and ICT Malaria Pf Tests in a Clinical Environment. J. Clin. Microbiol. 37: 1269-1273 [Abstract] [Full Text]  
• Fredricks, D. N., Relman, D. A. (1998). Improved Amplification of Microbial DNA from Blood Cultures by Removal of the PCR Inhibitor Sodium Polyanetholesulfonate. J. Clin. Microbiol. 36: 2810-2816 [Abstract] [Full Text]  
• Morandi, P.-A., Schockmel, G. A., Yerly, S., Burgisser, P., Erb, P., Matter, L., Sitavanc, R., Perrin, L. (1998). Detection of Human Immunodeficiency Virus Type 1 (HIV-1) RNA in Pools of Sera Negative for Antibodies to HIV-1 and HIV-2. J. Clin. Microbiol. 36: 1534-1538 [Abstract] [Full Text]  
• Dickover, R. E., Herman, S. A., Saddiq, K., Wafer, D., Dillon, M., Bryson, Y. J. (1998). Optimization of Specimen-Handling Procedures for Accurate Quantitation of Levels of Human Immunodeficiency Virus RNA in Plasma by Reverse Transcriptase PCR. J. Clin. Microbiol. 36: 1070-1073 [Abstract] [Full Text]  
• Parravicini, C., Lauri, E., Baldini, L., Neri, A., Poli, F., Sirchia, G., Moroni, M., Galli, M., Corbellino;, M., Masood, R., Zheng, T., Tulpule, A., Arora, N., Chatlynne, L., Handy, M., Whitman Jr., J., Kaplan, M., Dosik, M., Ablashi, D. V., Gill;, P. S., Whitby, D., Boshoff, C., Luppi, M., Torelli;, G., Cottoni, F., Uccini;, S., Brousset, P., Meggetto, F., Attal, M., Delsol;, G., Rettig, M. B., Sun, J. W. R., Berenson;, R. A. J. (1997). Kaposi's Sarcoma-Associated Herpesvirus Infection and Multiple Myeloma. Science 278: 1969-1973 [Full Text]  
• Natarajan, V, Plishka, R J, Scott, E W, Lane, H C, Salzman, N P (1994). An internally controlled virion PCR for the measurement of HIV-1 RNA in plasma.. Genome Res. 3: 346-350 [Abstract]  
• Burckhardt, J (1994). Amplification of DNA from whole blood.. Genome Res. 3: 239-243 [Abstract]  
• Nocton, J. J., Dressler, F., Rutledge, B. J., Rys, P. N., Persing, D. H., Steere, A. C. (1994). Detection of Borrelia burgdorferi DNA by Polymerase Chain Reaction in Synovial Fluid from Patients with Lyme Arthritis. NEJM 330: 229-234 [Abstract] [Full Text]  
• Poli, F, Cattaneo, R, Crespiatico, L, Nocco, A, Sirchia, G (1993). A rapid and simple method for reversing the inhibitory effect of heparin on PCR for HLA class II typing.. Genome Res. 2: 356-358