Screening potential corneal donors for HIV-1 by polymerase chain reaction and a colorimetric microwell hybridization assay.
OBJECTIVE
Current screening of potential corneal donors for human immunodeficiency virus type 1 (HIV-1) involves serologic detection of antibodies to the virus. However, this approach cannot detect infection during the seronegative window period of the disease. We therefore evaluated the polymerase chain reaction (PCR) assay for viral nucleic acid as a possible alternative to screening cadaveric blood for HIV-1.


METHODS
Blood specimens from cadavers diagnosed at autopsy with acquired immunodeficiency syndrome (AIDS) (n = 21), at high risk for HIV-1 infection (n = 47), and at no known risk (n = 350) were screened by PCR for HIV-1 proviral DNA and human leukocyte antigen (HLA)-DQ alpha sequences, and for HIV antibodies.


RESULTS
All AIDS group samples were seropositive; of these, 18 (86%) and 20 (95%) of 21 were positive for HIV by PCR of proteinase K- and Chelex-extracted pellets, respectively. The seropositive samples negative by PCR testing were shown to inhibit PCR amplification. Nine (19%) of 47 high-risk specimens were HIV-positive. The no-known-risk group yielded negative results. The overall sensitivities for PCR in the proteinase K- and Chelex-treated groups were 90% and 97%, respectively, compared with Western blot reactivity. If PCR-inhibitory samples and HLA-DQ alpha-negative samples had been eliminated, sensitivity would have been 100%. Specificity was 100% for each group.


CONCLUSIONS
Screening cadaveric blood by PCR may be feasible, but further refinement of the assay and blood specimen collection practices will be necessary for it to become routine. Future studies should focus on optimizing specimen procurement and preparation to reduce or eliminate specimens that inhibit PCR.
Essary
L R
LR
Department of Ophthalmology and Visual Sciences, Washington University School of Medicine, St. Louis, MO 63110, USA.
Kinard
S J
SJ
Butcher
A
A
Wang
H
H
Laycock
K A
KA
Donegan
E
E
McCreedy
B
B
Connell
S
S
Batchelor
J
J
Harris
J
J
Spadoro
J
J
Pepose
J S
JS
D015492:AIDS Serodiagnosis; D000163:Acquired Immunodeficiency Syndrome / Q000175:diagnosis; D015153:Blotting, Western; D003124:Colorimetry; D003315:Cornea; D017931:DNA Primers / Q000737:chemistry; D004279:DNA, Viral / Q000032:analysis; D004797:Enzyme-Linked Immunosorbent Assay; D015828:Eye Infections, Viral / Q000175:diagnosis*; D015483:HIV Antibodies / Q000032:analysis; D015658:HIV Infections / Q000175:diagnosis*; D006679:HIV Seropositivity / Q000175:diagnosis; D015497:HIV-1 / Q000235:genetics / Q000276:immunology; D006683:HLA-DQ Antigens / Q000032:analysis; D059848:HLA-DQ alpha-Chains; D006801:Humans; D008403:Mass Screening; D009693:Nucleic Acid Hybridization / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D014019:Tissue Donors
D017931:DNA Primers; D004279:DNA, Viral; D015483:HIV Antibodies; D006683:HLA-DQ Antigens; D059848:HLA-DQ alpha-Chains; C069050:HLA-DQA1 antigen