Abstract:
In recent years, it has become increasingly important to assure that transplanted organs and tissues are free of any contaminating agents that may result in disease and/or death to the recipient. Towards that effort, we are testing the efficacy of using PCR coupled with a non-radioactive microwell assay for the detection of HIV-1 in potential corneal donors. Blood is obtained from donors following expiration and prepared by a procedure that selectively lyses red blood cells, but leaves leukocytes intact. The leukocytes are extracted and then amplified by PCR in the presence of the primers, BIO-SK431/462. Following amplification, the reactions are analyzed by microwell hybridization using the SK102 capture probe. Cadaveric blood specimens are subject to unique problems not encountered in blood obtained from living patients. Because these samples are procured several hours post-mortem and are not necessarily from original venipuncture, they are often hemolyzed, contaminated with heparin, and hemodiluted. These suboptimal cadaveric samples therefore harbor potential PCR inhibitors and require that we test for sample integrity by amplifying for HLA-DQ-alpha. To date, 321 specimens from low risk, high risk, and known HIV-1 positive individuals have been examined by PCR. Of these samples, 95.4% were DQ-alpha positive. Of the DQ-alpha positive specimens, 4.6% were positive for HIV-1 by PCR, EIA, and WB. No low risk specimens yielded false positive results by PCR or serology. These results support the efficacy of using PCR to screen the blood of potential organ and tissue donors to reduce the potential risk of transmitting HIV-1 to the recipient.
Keywords: Cadaver *Corneal Transplantation Human HIV Seropositivity/BLOOD/*DIAGNOSIS/EPIDEMIOLOGY HIV-1/*ISOLATION & PURIF HLA-DQ Antigens/BLOOD *Nucleic Acid Hybridization Polymerase Chain Reaction/*METHODS Risk Factors *Tissue Donors ABSTRACT 930930
M9391149
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