Detection of pneumolysin in human urine using an immunosensor on screen-printed carbon electrodes

被引：13

作者：

Díaz-González, M ^{[1
]}

González-García, MB ^{[1
]}

Costa-García, A ^{[1
]}

机构：

[1] Univ Oviedo, Fac Quim, Dept Quim Fis & Analit, Oviedo 33006, Asturias, Spain

来源：

SENSORS AND ACTUATORS B-CHEMICAL | 2006年 / 113卷 / 02期

关键词：

immunosensor; alkaline phosphatase; 3-indoxyl phosphate; screen-printed carbon electrode; streptavidin-biotin reaction; pneumolysin;

D O I：

10.1016/j.snb.2005.04.019

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

A voltammetric immunosensor for pneumolysin (PLY), based on the electrochemical detection of indigo blue, produced by alkaline phosphatase (AP) when 3-indoxyl phosphate (3-IP) is used as enzymatic substrate, is prepared and evaluated for measuring this toxin in human urine samples. The single-use immunosensor is fabricated by deposition of biotinylated anti-PLY monoclonal antibodies onto pre-oxidised streptavidin coated screen-printed carbon electrodes (SPCEs). Rabbit polyclonal IgGs anti-PLY are used in combination with an anti-rabbit IgG alkaline phosphatase conjugate as detection antibodies. The estimated limit of detection of the purified recombinant toxin is 0.05 ng/ml in Tris-buffer and 0.12 ng/ml in undiluted urine samples, but the same linear range, between 0.25 and 5.0 ng/ml, is obtained in both cases. The immunosensor is also applied to detect pneumolysin in urine samples from two pneumococcal pneumonia patients. (c) 2005 Elsevier B.V. All rights reserved.

引用

页码：1005 / 1011

页数：7

共 38 条

[1] Flow amperometric detection of indigo for enzyme-linked immunosorbent assays with use of screen-printed electrodes [J].

Alvarez, MJB ;

Abedul, MTF ;

García, AC .

ANALYTICA CHIMICA ACTA, 2002, 462 (01) :31-37

[2] Rapid and reliable identification of Streptococcus pneumoniae isolates by pneumolysin-mediated agglutination [J].

Cima-Cabal, MD ;

Vázquez, F ;

de los Toyos, JR ;

Méndez, FJ .

JOURNAL OF CLINICAL MICROBIOLOGY, 1999, 37 (06) :1964-1966

[3] Immunodetection of pneumolysin in human urine by ELISA [J].

Cima-Cabal, MD ;

Méndez, FJ ;

Vázquez, F ;

Aranaz, C ;

Rodríguez-Alvarez, J ;

García-García, JM ;

Fleites, A ;

González-Río, JM ;

Molinos, L ;

de Miguel, D ;

de los Toyos, JR .

JOURNAL OF MICROBIOLOGICAL METHODS, 2003, 54 (01) :47-55

[4] A specific and ultrasensitive chemiluminescent sandwich ELISA test for the detection and quantitation of pneumolysin [J].

Cima-Cabal, MD ;

Méndez, FJ ;

Vázquez, F ;

Garcia-Suárez, MD ;

de los Toyos, JR .

JOURNAL OF IMMUNOASSAY & IMMUNOCHEMISTRY, 2001, 22 (02) :99-112

[5]

COONROD JD, 1976, J CLIN MICROBIOL, V4, P168

[6] Prospective study to determine clinical relevance of detection of pneumococcal DNA in sera of children by PCR [J].

Dagan, R ;

Shriker, O ;

Hazan, I ;

Leibovitz, E ;

Greenberg, D ;

Schlaeffer, F ;

Levy, R .

JOURNAL OF CLINICAL MICROBIOLOGY, 1998, 36 (03) :669-673

[7] Disposable amperometric immunosensor system for rabbit IgG using a conducting polymer modified screen-printed electrode [J].

Darain, F ;

Park, SU ;

Shim, YB .

BIOSENSORS & BIOELECTRONICS, 2003, 18 (5-6) :773-780

[8] Functional analysis of pneumolysin by use of monoclonal antibodies [J].

delosToyos, JR ;

Mendez, FJ ;

Aparicio, JF ;

Vazquez, F ;

Suarez, MD ;

Fleites, A ;

Hardisson, C ;

Morgan, PJ ;

Andrew, PW ;

Mitchell, TJ .

INFECTION AND IMMUNITY, 1996, 64 (02) :480-484

[9] Development of an immunosensor for the determination of rabbit IgG using streptavidin modified screen-printed carbon electrodes [J].

Díaz-González, M ;

Hernández-Santos, D ;

González-García, MB ;

Costa-García, A .

TALANTA, 2005, 65 (02) :565-573

[10]

Díaz-González M, 2002, ELECTROANAL, V14, P665, DOI 10.1002/1521-4109(200205)14:10<665::AID-ELAN665>3.0.CO

← 1 2 3 4 →