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 Yourno, J Articles by Conroy, J Search for Related Content PubMed PubMed Citation Articles by Yourno, J Articles by Conroy, J

A novel polymerase chain reaction method for detection of human immunodeficiency virus in dried blood spots on filter paper.

New York State Department of Health, Wadsworth Center for Laboratory and Research, Empire State Plaza, Albany, New York 12201-0509.

ABSTRACT

A method for detection of proviral human immunodeficiency virus DNA in dried blood spots on filter paper by direct polymerase chain reaction (PCR) has been developed. To develop the method, a standard system was used which was prepared from cells each containing a single integrated provirus and titrated with normal donor blood. This rapid procedure provides virtually quantitative yields of nuclear DNA and exploits most of the standard methodology described for blood specimens. A nested PCR using SK38-SK39 gag as the internal primer pair was also designed; this PCR detected a single copy of provirus per filter at near theoretical frequency with SK19 probe. The utility of the procedure was demonstrated with clinical specimens. Blood spot filters from human immunodeficiency virus-infected and uninfected individuals were readily and unequivocally discriminated. The method is designed for ultimate use with large (1.5-ml) sample preparation tubes that are compatible as PCR tubes with thermal cyclers. This will permit convenient, direct single-tube PCR of dried blood specimens on filters. It should be adaptable to analysis of dried blood spots for a variety of infectious or genetic diseases.

This article has been cited by other articles:

• Nozawa, N., Koyano, S., Yamamoto, Y., Inami, Y., Kurane, I., Inoue, N. (2007). Real-Time PCR Assay Using Specimens on Filter Disks as a Template for Detection of Cytomegalovirus in Urine. J. Clin. Microbiol. 45: 1305-1307 [Abstract] [Full Text]  
• NOUHIN, J., NGUYEN, M. (2006). EVALUATION OF A BOOSTED-P24 ANTIGEN ASSAY FOR THE EARLY DIAGNOSIS OF PEDIATRIC HIV-1 INFECTION IN CAMBODIA. Am J Trop Med Hyg 75: 1103-1105 [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]  
• Auewarakul, P., Sangsiriwut, K., Suwanagool, S., Wasi, C. (2001). Target Cell Populations of Human Immunodeficiency Virus Type 1 in Peripheral Blood Lymphocytes with Different Chemokine Receptors at Various Stages of Disease Progression. J. Virol. 75: 6384-6391 [Abstract] [Full Text]  
• Panteleeff, D. D., John, G., Nduati, R., Mbori-Ngacha, D., Richardson, B., Kreiss, J., Overbaugh, J. (1999). Rapid Method for Screening Dried Blood Samples on Filter Paper for Human Immunodeficiency Virus Type 1 DNA. J. Clin. Microbiol. 37: 350-353 [Abstract] [Full Text]