Development of PCR-ELISA for Detection of HIV-1 in Biomedical Products.
- Author:
Jung Hyun SHIM
1
;
Jong Wan KIM
;
Kyung Ae LEE
;
Dur Han KWON
;
Pyung Keun MYUNG
;
Jae Ok KIM
;
Seok Ho LEE
;
Sue Nie PARK
;
Do Young YOON
Author Information
1. Laboratory of Cell Biology, Korea Rearch Institute of Bioscience & Biotechnology, Daejeon, Korea. dyyoon@kribb.re.kr
- Publication Type:Original Article
- Keywords:
HIV-1;
PCR-ELISA
- MeSH:
Biological Products;
Culture Media;
Enzyme-Linked Immunosorbent Assay;
HIV-1*;
Limit of Detection;
Plasmids;
Polymerase Chain Reaction;
Sepharose;
Viral Vaccines
- From:The Korean Journal of Laboratory Medicine
2004;24(1):60-66
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
BACKGROUND: Biomedical products such as viral vaccines can be contaminated with hazardous viruses during manufacturing processes and storage, thus causing harmful side effects. To assure the safety of biomedical products, highly effective and sensitive methods should be available to detect contaminating viruses. In this study, we performed recovery tests to determine the limit of detection of HIV-1. METHODS: An HIV-1 plasmid preparation was serially diluted and spiked into various culture media (DMEM, RPMI-1640, IMDM, GICM, and SDM) containing 10% fetal bovine serum (FBS). The HIV-1 plasmid was detected by PCR alone or a combination of PCR and ELISA (PCR/ELISA). RESULTS: When spiked into DMEM, RPMI, and IMDM, less than 4x10(-2) ng of HIV-1 plasmid was not detectable as HIV-1 PCR products in agarose gel. Intra- and inter-assays (n=6) showed that the PCR-ELISA system could detect PCR products diluted as much as 1, 875 times from HIV-1 plasmid serially spiked in various media. CONCLUSIONS: The PCR/ELISA system can be useful for the detection of trace amounts of hazardous viruses which may be present as contaminants in biological products.
