Temperature effect on the sensitivity of ELISA, PA and WB to detect anti-HIV-1 antibody and infectivity of HIV-1.
BACKGROUND
This study is designed to resolve the problem of whether temperature or freeze/thaw cycle will have any impact on the sensitivity for detection of anti-HIV-1 antibody by particle agglutination (PA), enzyme-linked immunosorbent assay (ELISA) and western blotting (WB). To reduce potential risk for laboratory personnel exposed to HIV-infection, it will be useful to determine the temperature effect on HIV infectivity.


METHODS
Testing sera were incubated at different temperatures or treated with several cycles of freeze and thaw. PA, ELISA and WB were used to detect anti-HIV-antibodies, whereas syncytia formation assay and polymerase chain reaction (PCR) were applied to detect HIV-infection.


RESULTS
The data showed that certain temperature points (no treatment, 25 degrees C for 1 hr, 2 hrs and 4 hrs, 37 degrees C for 30 minutes and 60 minutes, 56 degrees C for 30 minutes and 60 minutes, 65 degrees C for 15 minutes and 30 minutes) had no impact on the testing results of ELISA, PA and WB in detection of anti-HIV-1 antibody. In addition, testing results of 50 normal human serum samples which had been heated to 56 degrees C for 30 minutes were still negative by ELISA and PA. Only the samples incubated at 65 degrees C for 60 minutes had slight differences in results. Freeze and thaw treatments of the serum did not alter anti-HIV testing results, either. Treatments of supernatant of HTLV-IIIB culture at 56 degrees C for 30 minutes and 60 minutes, 65 degrees C for 15 minutes and 30 minutes could eliminate the syncytia formation caused by HIV-infection. Further analysis of the samples by PCR was able to detect HIV-specific sequences in all the treatments.


CONCLUSIONS
Anti-HIV antibody is quite stable in serum, even when it is pre-heated to 56 degrees C for 30 minutes. Freeze and thaw treatment of serum samples up to seven cycles did not change the results, either. In addition, to minimize the potential risk of laboratory personnel exposed to HIV infection, pre-treatment of serum samples with heat at 56 degrees C for 30 minutes or 60 minutes can reduce HIV infectivity. However, laboratories still must emphasize the importance of universal precautions rather than heat-inactivation of serum to prevent occupational transmission of HIV.
Wang
G R
GR
Department of Medicine, Taipei Medical College, Taiwan, R.O.C.
Yang
J Y
JY
Lin
T L
TL
Chen
H Y
HY
Horng
C B
CB
D000372:Agglutination Tests; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D015483:HIV Antibodies / Q000097:blood*; D015497:HIV-1 / Q000276:immunology* / Q000472:pathogenicity; D006801:Humans; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D013696:Temperature
D015483:HIV Antibodies