Chemiluminescent methods for detecting and quantitating enzyme activity

Kricka, Larry J.; Voyta, John C.; Bronstein, Irena

doi:10.1016/s0076-6879(00)05500-2

Cited by 54 publications

(47 citation statements)

References 64 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…It is not possible to distinguish the relative contributions of these two parameters to the increased sensitivity of the ZstatFlu-II test. However, it is commonly accepted by chemistry analyzers that chemiluminescence is more sensitive than enzyme immunoassays (14). This suggests that the detection method contributed to the increased sensitivity of the ZstatFlu-II test (1, 4).…”

Section: Discussionmentioning

confidence: 99%

“…This approach for influenza virus detection is similar to that of the previously reported ZstatFlu test, which employed a chromogenic substrate specific for influenza virus neuraminidase (18,20). In chemistry analyzers, chemiluminescence is generally accepted as being more sensitive than enzyme immunoassays (14). Chemiluminescence provides an opportunity for increasing the sensitivity of rapid tests (1,4).…”

mentioning

confidence: 99%

See 1 more Smart Citation

Clinical Evaluation of the ZstatFlu-II Test: a Chemiluminescent Rapid Diagnostic Test for Influenza Virus

et al. 2002

View full text Add to dashboard Cite

Exploiting the high sensitivity of the chemiluminescence phenomenon, an accurate and sensitive point-ofcare test, called the ZstatFlu-II test (ZymeTx, Inc., Oklahoma City, Okla.), was developed to detect influenza virus infections. The ZstatFlu-II test takes 20 min and requires approximately 2 min of "hands-on" time for operational steps. The ZstatFlu-II test does not distinguish between infections with influenza virus types A and B. ZstatFlu-II test results are printed on Polaroid High-Speed Detector Film, allowing test results to be archived. A prototype version of the ZstatFlu-II test was evaluated during the 2000-to-2001 flu season with 300 nasal aspirate specimens from children at a pediatric hospital. Compared to culture, the ZstatFlu-II test had 88% sensitivity and 92% specificity. The Directigen test had a sensitivity of 75% and a specificity of 93%. The sensitivity of the ZstatFlu-II test was significantly higher than that of the Directigen test (P < 0.0574).The annual economic cost of influenza disease in the United States has been estimated at $3 billion to $5 billion (22). The Centers for Disease Control and Prevention reported in 2001 that influenza was associated with about 20,000 deaths nationwide and more than 100,000 hospitalizations (http://www.cdc .gov/ncidod/diseases/flu/fluinfo.htm). Worldwide estimates were considerably higher (16,17,24). Sensitive, specific, and rapid tests for influenza will greatly improve patient health care and reduce costs so that only "flu-positive" patients receive the recently approved antiviral treatments (9, 21). Rapid diagnostics for influenza will also prevent the misuse of antibiotics to "treat" the flu.Influenza disease is caused by influenza virus types A and B. Influenza virus types A and B are Orthomyxoviridae, characterized by the presence of an envelope penetrated by glycoprotein spikes with hemagglutinating and neuraminidase activities. Influenza virus also contains matrix protein, nucleoprotein, and three proteins with polymerase activity and a segmented negative-strand RNA genome (6, 19). These viruses are responsible for winter epidemics of respiratory illness in which the rates of infection are highest among children (5). The shared cardinal sign of fever without localization makes differentiation of influenza from sepsis necessary for proper patient management. Delay in this differentiation could result in unnecessary laboratory testing and treatment for possible bacterial "sepsis" with unnecessary antibiotics (25).Three diagnostic methods for respiratory secretions are in common use. Culture, both shell vial and tube, takes several days. Direct or indirect immunofluorescence assays on exfoliated nasal pharyngeal cells could be done in a few hours but require a high level of expertise (10). Finally, rapid, point-ofcare tests are available to detect the influenza virus (3,12,20,23). All of these tests use an enzyme immunoassay directed at antigens of the viruses (3, 12, 23), with the exception of the ZstatFlu test, which detects influenza virus neur...

show abstract

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

Clinical Evaluation of the ZstatFlu-II Test: a Chemiluminescent Rapid Diagnostic Test for Influenza Virus

et al. 2002

View full text Add to dashboard Cite

show abstract

“…The enhanced chemiluminescence (CL) of the luminolhydrogen peroxide (H 2 O 2 )-horseradish peroxidase (HRP) system using phenolic enhancers has been widely applied to the detection of HRP labels in an enzyme immunoassay, enzymatically generated H 2 O 2 and other analytes (1)(2)(3)(4)(5)(6)(7)(8). We recently reported a novel enhancer, 2-(4-hydroxyphenyl)-4,5-diphenylimidazole (HDI), for the luminol-H 2 O 2 -HRP system (9), and applied it to the development of a lauric acid ester of HDI (HDIlaurate) as a proenhancer type substrate for lipase (triacylglycerol lipase; EC.3.1.1.3) (5).…”

Section: Introductionmentioning

confidence: 99%

Study on immunocapture–chemiluminescence assay of lipase activity in a biological sample

et al. 2005

View full text Add to dashboard Cite

A new approach for the determination of lipase (triacylglycerol lipase, EC.3.1.1.3) activity in a biological sample was investigated by combining an immunocapture technique with a chemiluminescence (CL) assay method in order to eliminate interference with CL detection. The proposed method consists of an immunocapture step to trap lipase and a subsequent step for CL detection of the activity of the captured lipase. The CL detection is based on the luminol-hydrogen peroxide (H(2)O(2))-horseradish peroxidase (HRP) reaction and utilizes a proenhancer substrate [a lauric acid ester of 2-(4-hydroxyphenyl)-4,5-diphenylimidazole (HDI)] which liberates an active enhancer, HDI, by enzymatic hydrolysis. A polyclonal antibody prepared with porcine pancreas lipase was used for the immunocapture. The proposed immunocapture-CL method effectively eliminated the interference with the CL reaction from biological components and enabled the determination of spiked porcine pancreas lipase activity in serum samples in the range 0.41-1.1 U(HDI) (1 U(HDI) corresponds to the amount which liberates 1 pmol HDI/min at 37 degrees C from the substrate). The method was further applied to the assay of the activity for human pancreas lipase in serum and the results showed good correlation (r = 0.871) with those by the conventional colorimetric method.

show abstract

“…The acyl group is transferred to the primary amine, resulting in a product that is covalently cross-linked by an isopeptide bond (1). Chemiluminescence (CL) is a powerful technique for probing a wide range of biological activities, including highly sensitive enzymatic assays (2)(3)(4)(5). We previously showed that N-(4-aminobutyl)-N-ethylisoluminol (ABEI) was a sensitive CL substrate for tissue transglutaminase and blood coagulation factor XIIIa (6,7).…”

Section: Introductionmentioning

confidence: 99%

Thiols enhance the sensitivity of luminescent assays for non‐cross‐linked and covalently cross‐linked aminophthalhydrazides

Achyuthan¹

2001

Luminescence

View full text Add to dashboard Cite

Transglutaminases catalyse an acyl-transfer reaction between the gamma-carboxamide of a protein or peptide-bound glutamine (P-CH2-gammaCH2-CO-1NH2) and the primary amino group of mono- or polyamines (R-2NH2), covalently cross-linking the reactants by an isopeptide bond: P-CH2-gammaCH2-CO-1NH2 + R-2NH2 --> P-CH2-gammaCH2-CO-2NH-R + 1NH3. We reported that N-(4-aminobutyl)-N-ethylisoluminol (ABEI) was a chemiluminescent (CL) amine substrate for transglutaminases. We now identified N-(6-aminohexyl)-N-ethylisoluminol (AHEI) as a second, less reactive, transglutaminase substrate. A structure-based explanation is offered for the lower reactivity of AHEI. Optimum CL from non-cross-linked or cross-linked ABEI or AHEI was elicited in the presence of 10 mmol/L dithiothreitol, by oxidizing with a mixture of 20 mmol/L potassium ferricyanide and 10 mmol/L hydrogen peroxide in 100 mmol/L NaOH. The limits of detection and quantitation for non-cross-linked aminophthalhydrazides obtained in this system were: 20 fmol and 60 fmol for ABEI and 10 fmol and 30 fmol for AHEI. These values represented a 500-800-fold improved sensitivity. Delayed peak CL and CL decay in the presence of dithiothreitol contributed to improving the sensitivity. The data could be useful for improving the immunoassays for aminophthalhydrazides and facilitate the development of high throughput assays for transglutaminases.

show abstract

Chemiluminescent methods for detecting and quantitating enzyme activity

Cited by 54 publications

References 64 publications

Clinical Evaluation of the ZstatFlu-II Test: a Chemiluminescent Rapid Diagnostic Test for Influenza Virus

Clinical Evaluation of the ZstatFlu-II Test: a Chemiluminescent Rapid Diagnostic Test for Influenza Virus

Study on immunocapture–chemiluminescence assay of lipase activity in a biological sample

Thiols enhance the sensitivity of luminescent assays for non‐cross‐linked and covalently cross‐linked aminophthalhydrazides

Contact Info

Product

Resources

About