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 Cassol, S Articles by O'Shaughnessy, M V Search for Related Content PubMed PubMed Citation Articles by Cassol, S Articles by O'Shaughnessy, M V

Stability of dried blood spot specimens for detection of human immunodeficiency virus DNA by polymerase chain reaction.

Centre for Excellence in HIV/AIDS, University of British Columbia, Vancouver, Canada.

ABSTRACT

Blood sampling on filter paper has many advantages for the detection of perinatal human immunodeficiency virus (HIV) infection by the polymerase chain reaction (PCR). However, if the method is to be widely used, an assessment of its performance under field conditions is required. To simulate conditions in the field, 50-microliters aliquots of whole blood containing low levels of HIV proviral DNA (4 to 1,024 copies per 100,000 nucleated cells) were spotted onto filter paper; dried; and subjected to heat, humidity, and prolonged storage at room temperature. After exposure, the DNA was recovered and amplified with primers to human leukocyte antigen DQ alpha- and HIV-specific sequences. Treatment at 37 degrees C and 60% humidity for 7 days, storage for 12 weeks at 22 degrees C, and freeze-thawing twice had no adverse effect on PCR reactivity when compared with the results obtained with reference spots stored at -20 degrees C. The lower limits of HIV detection in all tests ranged from 4 to 16 HIV copies per 100,000 cells. Fixation in 70% ethanol improved the amplification of low levels of HIV DNA and reduced biohazard risks. These findings suggest that dried blood spots will provide a powerful new resource for testing for HIV by PCR, especially in remote areas where refrigeration and immediate sample processing are unavailable.

This article has been cited by other articles:

• Ayele, W., Schuurman, R., Messele, T., Dorigo-Zetsma, W., Mengistu, Y., Goudsmit, J., Paxton, W. A., de Baar, M. P., Pollakis, G. (2007). Use of Dried Spots of Whole Blood, Plasma, and Mother's Milk Collected on Filter Paper for Measurement of Human Immunodeficiency Virus Type 1 Burden. J. Clin. Microbiol. 45: 891-896 [Abstract] [Full Text]  
• Patton, J. C., Akkers, E., Coovadia, A. H., Meyers, T. M., Stevens, W. S., Sherman, G. G. (2007). Evaluation of Dried Whole Blood Spots Obtained by Heel or Finger Stick as an Alternative to Venous Blood for Diagnosis of Human Immunodeficiency Virus Type 1 Infection in Vertically Exposed Infants in the Routine Diagnostic Laboratory. CVI 14: 201-203 [Abstract] [Full Text]  
• Luo, W., Yang, H., Rathbun, K., Pau, C.-P., Ou, C.-Y. (2005). Detection of Human Immunodeficiency Virus Type 1 DNA in Dried Blood Spots by a Duplex Real-Time PCR Assay. J. Clin. Microbiol. 43: 1851-1857 [Abstract] [Full Text]  
• Li, C.-C., Beck, I. A., Seidel, K. D., Frenkel, L. M. (2004). Persistence of Human Immunodeficiency Virus Type 1 Subtype B DNA in Dried-Blood Samples on FTA Filter Paper. J. Clin. Microbiol. 42: 3847-3849 [Abstract] [Full Text]  
• Brambilla, D., Jennings, C., Aldrovandi, G., Bremer, J., Comeau, A. M., Cassol, S. A., Dickover, R., Jackson, J. B., Pitt, J., Sullivan, J. L., Butcher, A., Grosso, L., Reichelderfer, P., Fiscus, S. A. (2003). Multicenter Evaluation of Use of Dried Blood and Plasma Spot Specimens in Quantitative Assays for Human Immunodeficiency Virus RNA: Measurement, Precision, and RNA Stability. J. Clin. Microbiol. 41: 1888-1893 [Abstract] [Full Text]  
• Wang, C.-Y. J., Giambrone, J. J., Smith, B. F. (2002). Detection of Duck Hepatitis B Virus DNA on Filter Paper by PCR and SYBR Green Dye-Based Quantitative PCR. J. Clin. Microbiol. 40: 2584-2590 [Abstract] [Full Text]  
• Tetali, S., Lee, E. M., Kaplan, M. H., Romano, J. W., Ginocchio, C. C. (2001). Chemokine Receptor CCR5 {Delta}32 Genetic Analysis Using Multiple Specimen Types and the NucliSens Basic Kit. CVI 8: 965-971 [Abstract] [Full Text]  
• Beck, I. A., Drennan, K. D., Melvin, A. J., Mohan, K. M., Herz, A. M., Alarcón, J., Piscoya, J., Velázquez, C., Frenkel, L. M. (2001). Simple, Sensitive, and Specific Detection of Human Immunodeficiency Virus Type 1 Subtype B DNA in Dried Blood Samples for Diagnosis in Infants in the Field. J. Clin. Microbiol. 39: 29-33 [Abstract] [Full Text]  
• Smyth, A, Tong, C Y W, Carty, H, Hart, C A (1997). Impact of HIV on mortality from acute lower respiratory tract infection in rural Zambia. Arch. Dis. Child. 77: 227-230 [Abstract] [Full Text]