DEVELOPMENT OF A SENSITIVE ELISA FOR HIV-1 P24 ANTIGEN USING A FLUOROGENIC SUBSTRATE FOR MONITORING HIV-1 REPLICATION INVITRO

被引：5

作者：

CHARGELEGUE, D

OTOOLE, CM

机构：

[1] Department of Medical Microbiology, London Hospital Medical College, London

来源：

JOURNAL OF VIROLOGICAL METHODS | 1992年 / 38卷 / 03期

关键词：

HIV-1; P24; ANTIGEN; ELISA; FLUORESCENCE; BETA-GALACTOSIDASE;

D O I：

10.1016/0166-0934(92)90077-Q

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A two-site sandwich fluorescent-ELISA was optimized for the detection of HIV-1 p24 antigen produced by lymphoid cells infected with HIV-1 in vitro. To improve the sensitivity of the ELISA, a combination of streptavidin-beta-galactosidase and a fluorogenic substrate (4-methylumbelliferyl-beta-D-galactopyranoside) was employed for the enzymatic detection stage. Using recombinant p24 as standard antigen, a two-step assay detected as little as 0.7 pg/ml (3.10(-14) M) with an upper limit of 10000 pg/ml. This detection range (approx. 50 70-times greater than ELISAs using a chromogenic detection) permitted an accurate and straightforward quantitation of p24 in culture supernatants. Overall, the fluorescent-ELISA had increased detectability, sensitivity and efficiency over existing ELISAs for HIV-1 p24.

引用

页码：323 / 332

页数：10

共 16 条

[1] ISOLATION OF A T-LYMPHOTROPIC RETROVIRUS FROM A PATIENT AT RISK FOR ACQUIRED IMMUNE-DEFICIENCY SYNDROME (AIDS) [J].