Flow microsphere immunoassay-based method of virus quantitation

被引：14

作者：

Samoylova, TI ^{[1
]}

Smith, BF ^{[1
]}

机构：

[1] Auburn Univ, Coll Vet Med, Scott Ritchey Res Ctr, Auburn, AL 36849 USA

来源：

BIOTECHNIQUES | 1999年 / 27卷 / 02期

关键词：

D O I：

10.2144/99272rr03

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A sensitive assay for adenovirus quantitation in vitro was developed using the flow microsphere immunoassay (FMIA) approach. Polystyrene microspheres were covalently coated with purified anti-adenoviral antibodies and incubated with virus-containing samples. After incubation, the samples were stained with DNA-specific fluorescent dyes, and microsphere-associated fluorescence was quantitated with a flow cytometer: The adsorption of virus to microspheres was examined under different experimental conditions. The flow cytometric assay was determined to be as accurate in detecting adenovirus as titering on 293 cells. The proposed method can be used to quantify virus in viral stock and in biological samples.

引用

页码：356 / 361

页数：6

共 16 条

[1] A flow cytometric immunoassay for β2-microglobulin in whole blood [J].

Bishop, JE ;

Davis, KA .

JOURNAL OF IMMUNOLOGICAL METHODS, 1997, 210 (01) :79-87

[2] ADENOVIRUS HEXON MONOCLONAL-ANTIBODY THAT IS GROUP-SPECIFIC AND POTENTIALLY USEFUL AS A DIAGNOSTIC REAGENT [J].

CEPKO, CL ;

WHETSTONE, CA ;

SHARP, PA .

JOURNAL OF CLINICAL MICROBIOLOGY, 1983, 17 (02) :360-364

[3] Retroviral infection is limited by Brownian motion [J].

Chuck, AS ;

Clarke, MF ;

Palsson, BO .

HUMAN GENE THERAPY, 1996, 7 (13) :1527-1534

[4] Consistent and high rates of gene transfer can be obtained using flow-through transduction over a wide range of retroviral titers [J].

Chuck, AS ;

Palsson, BO .

HUMAN GENE THERAPY, 1996, 7 (06) :743-750

[5]

HAUGLAND RP, 1994, METHOD CELL BIOL, V42, P641

[6] SOLID-PHASE IMMUNOASSAY USING A FLOW CYTOMETER - QUANTITATIVE AND QUALITATIVE DETERMINATION OF PROTEIN ANTIGENS AND A HAPTEN [J].

KIM, K ;

HAN, MY ;

YOON, DY ;

CHO, B ;

CHOI, MJ ;

CHOE, IS ;

CHUNG, TW .

IMMUNOLOGY LETTERS, 1992, 31 (03) :267-272

[7]

Lennette DA, 1995, DIAGNOSTIC PROCEDURE, P3

[8] POLYPEPTIDES OF ADENOVIRUS .I. EVIDENCE FOR MULTIPLE PROTEIN COMPONENTS IN VIRION AND A COMPARISON OF TYPES 2 7A AND 12 [J].

MAIZEL, JV ;

WHITE, DO ;

SCHARFF, MD .

VIROLOGY, 1968, 36 (01) :115-+

[9]

McHugh TM, 1997, CYTOMETRY, V29, P106

[10] SIMULTANEOUS DETECTION OF ANTIBODIES TO CYTOMEGALO-VIRUS AND HERPES-SIMPLEX VIRUS BY USING FLOW-CYTOMETRY AND A MICROSPHERE-BASED FLUORESCENCE IMMUNOASSAY [J].

MCHUGH, TM ;

MINER, RC ;

LOGAN, LH ;

STITES, DP .

JOURNAL OF CLINICAL MICROBIOLOGY, 1988, 26 (10) :1957-1961

← 1 2 →