Affiliation:
1. 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.
Publisher
American Society for Microbiology
Reference32 articles.
1. Effect of fixation on the amplification of nucleic acids from paraffin-embedded material by the polymerase chain reaction;Ben-Ezra J.;J. Histochem. Cytochem.,1991
2. Use of dried blood spot specimens in the detection of human immunodeficiency virus type 1 by the polymerase chain reaction;Cassol S.;J. Clin. Microbiol.,1991
3. Diagnosis of vertical HIV-1 transmission using the polymerase chain reaction and dried blood spot specimens;Cassol S. A.;J. Acquired Immune Defic. Syndr.,1992
4. Centers for Disease Control. 1990. HIV/AIDS surveillance report p. 1-18. Centers for Disease Control Atlanta.
5. Current and future dimensions of the HIV and AIDS epidemic in women and children;Chin J.;Lancet,1990
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