Antibody development and enzyme-linked immunosorbent assay for the protein marker lactate dehydrogenase to determine safe cooking end-point temperatures of turkey rolls

1992 ◽  
Vol 40 (9) ◽  
pp. 1671-1676 ◽  
Author(s):  
Cheng Hsin. Wang ◽  
Mohamed M. Abouzied ◽  
James J. Pestka ◽  
Denise M. Smith
Keyword(s):  
Lactate Dehydrogenase ◽  
Immunosorbent Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Protein Marker ◽  
End Point

Lactate Dehydrogenase Polyclonal Antibody Sandwich ELISA for Determination of Endpoint Heating Temperatures of Ground Beef

1996 ◽  
Vol 59 (1) ◽  
pp. 51-55 ◽  
Author(s):  
CHENG-HSIN WANG ◽  
MOHAMED M. ABOUZIED ◽  
JAMES J. PESTKA ◽  
DENISE M. SMITH
Keyword(s):  
Lactate Dehydrogenase ◽  
Polyclonal Antibody ◽  
Heating Temperature ◽  
Polyclonal Antibodies ◽  
Sandwich Elisa ◽  
Ground Beef ◽  
Enzyme Linked Immunosorbent Assay ◽  
Protein Marker ◽  
Beef Patties

An enzyme-linked immunosorbent assay (ELISA) for the protein marker lactate dehydrogenase (LDH) was developed to determine endpoint heating temperature (EPT) of ground beef. Ground beef was placed in 10 by 75 mm tubes and heated to temperatures between 62 and 74°C at 2°C intervals. Electrophoresis and immunoblotting of meat extracts indicated a decrease in the intensity of the LDH band as temperature was increased. Polyclonal antibodies (PAb) against bovine muscle LDH were produced and a sandwich ELISA devised using biotinylated PAb as the detecting antibody. The LDH content decreased (P < 0.05) in ground beef heated between 66 and 74°C and was less than 4 μg/g of meat at the recommended minimum endpoint of 69°C. The ELISA was used to estimate the EPT of commercially cooked beef patties.

scholarly journals Improved Assessment of Plasmodium vivax Response to Antimalarial Drugs by a Colorimetric Double-Site Plasmodium Lactate Dehydrogenase Antigen Capture Enzyme-Linked Immunosorbent Assay

2007 ◽  
Vol 51 (6) ◽  
pp. 2112-2116 ◽  
Author(s):  
Pierre Druilhe ◽  
Philippe Brasseur ◽  
Catherine Blanc ◽  
Michael Makler
Keyword(s):  
Lactate Dehydrogenase ◽  
Plasmodium Vivax ◽  
Immunosorbent Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Response Curves ◽  
In Vitro Susceptibility ◽  
Poor Growth ◽  
Antigen Capture

ABSTRACT The occurrence of Plasmodium vivax resistance to chloroquine has been reported in several countries of Asia and South America. However, the resistance of P. vivax is insufficiently documented for three reasons: it has received far less attention than P. falciparum; in vivo investigations are handicapped by the existence of hypnozoites, which make it difficult to distinguish between recrudescences due to drug failure and relapses due to dormant forms in the liver; and in vitro studies are greatly limited by the poor growth of P. vivax. We report on the adaptation to P. vivax of a colorimetric double-site Plasmodium lactate dehydrogenase antigen capture enzyme-linked immunosorbent assay previously developed for P. falciparum. The assay proved remarkably sensitive, as under optimal conditions it could detect P. vivax parasitemia levels as low as 10−8. The technique, which relies on the detection of protein synthesis by the parasite, yielded steep drug-response curves, leading to the precise determination of the 50% inhibitory concentrations for a high proportion of isolates. Chloroquine-resistant parasites were identified in an area where this phenomenon had been documented by in vivo methods. Thus, the results indicate that the in vitro susceptibility of P. vivax can now be monitored easily and efficiently. The data suggest that the threshold of resistance is similar to that of P. falciparum, i.e., in the range of 100 nM for chloroquine and 15 nM for pyronaridine. However, further studies are required to precisely define the cutoff for resistance and the sensitivity to each drug.

Sensitive microplate enzyme-linked immunosorbent assay for detection of antibodies against the scrub typhus rickettsia, Rickettsia tsutsugamushi

1979 ◽  
Vol 9 (1) ◽  
pp. 38-48 ◽  
Author(s):  
G A Dasch ◽  
S Halle ◽  
A L Bourgeois
Keyword(s):  
Optical Density ◽  
Immunosorbent Assay ◽  
Scrub Typhus ◽  
Fluorescent Antibody ◽  
Antibody Test ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Igg Antibody ◽  
Serum Dilution ◽  
End Point

A microtiter enzyme-linked immunosorbent assay (ELISA) has been developed for the titration of antibodies against scrub typhus in human and animal sera. Scrub typhus rickettsiae were grown in monolayers of irradiated mouse LM3 cells and separated from host cell materials by differential centrifugation, filtration through a glass filter (AP-20, Millipore Corp.), and isopycnic banding in Renografin density gradients. The scrub typhus ELISA antigens were obtained from the purified viable rickettsiae by French pressure cell disruption and addition of 0.2% Formalin to the soluble extract. Antisera prepared in rabbits against the prototype Karp, the Kato, and the Gilliam strains of scrub typhus were used to standardize the ELISA and to compare its sensitivity and specificity to that of the indirect fluorescent antibody test (IFA). ELISA titers were measured as the greatest serum dilution showing an optical density 0.25 above controls or by the optical density achieved at a fixed serum dilution. The IFA and ELISA end point titers were quite similar, and all three measures of titer had comparable specificity for the strains of scrub typhus. No cross-reactions between the typhus and scrub typhus wera were observed by ELISA. Both the immunoglobulin M (IgM) and IgG antibody titers of 12 sequential sera from four patients with scrub typhus were obtained by IFA and ELISA. The IFA and ELISA end point titers for IgM and IgG had correlation coefficients of 0.91 and 0.97, respectively, whereas the ELISA optical density values at a serum dilution of 1:100 had slightly lower correlations with IFA titers (0.80 and 0.94). Early rising IgM titers followed by rising IgG titers were demonstrated by ELISA in three patients with primary scrub typhus infections, whereas the IgG response predominated in a patient with a reinfection. It is concluded that the ELISA for scrub typhus is a very satisfactory alternative to the IFA test.

Lactate Dehydrogenase as Safe Endpoint Cooking Indicator in Poultry Breast Rolls: Development of Monoclonal Antibodies and Application to Sandwich Enzyme-Linked Immunosorbent Assay (ELISA)

1993 ◽  
Vol 56 (2) ◽  
pp. 120-124 ◽  
Author(s):  
MOHAMED M. ABOUZIED ◽  
CHENG HSING WANG ◽  
JAMES J. PESTKA ◽  
DENISE M. SMITH
Keyword(s):  
Monoclonal Antibodies ◽  
Lactate Dehydrogenase ◽  
Immunosorbent Assay ◽  
Polyclonal Antibodies ◽  
Sandwich Elisa ◽  
Enzyme Linked Immunosorbent Assay ◽  
Department Of Agriculture ◽  
Poultry Products ◽  
Chicken Muscle ◽  
Assay Detection

A sandwich enzyme-linked immunosorbent assay (ELISA) was developed to detect lactate dehydrogenase (LDH) as a marker protein for verifying endpoint cooking of uncured poultry products. Monoclonal antibodies were prepared against chicken muscle LDH and used with rabbit polyclonal antibodies developed against turkey or chicken muscle LDH for capture and detection in the assay, respectively. Minimum assay detection limits for turkey and chicken muscle LDH were 1 ng/ml. Turkey and chicken muscle LDH, but not LDH from other species cross reacted in the ELISA. The ELISA was further verified using extracts of turkey breast rolls processed to internal temperatures between 68.3 and 72.1°C. The LDH content of extracts diluted 3- to 6-fold was below 15 ng/ml for turkey rolls processed to 70.9 and 72.1°C. At a 6-fold dilution, LDH content of extracts from rolls processed to 69.7°C was approximately 10 times greater than those processed to 70.9°C. A survey of market precooked poultry products indicated assay validity with precooked turkey roast, but not turkey hams with maximum internal temperature requirements of 68.3°C. Results suggested the sandwich ELISA should be applicable for determining whether turkey breast rolls are processed to the required U.S. Department of Agriculture endpoint temperature of 71.1°C.

Rapid Enzyme-Linked Immunosorbent Assay (ELISA) with a visual end-point for detecting opiate narcotics in urine

The Analyst ◽  
1987 ◽  
Vol 112 (5) ◽  
pp. 687 ◽  
Author(s):  
David Laurie ◽  
Andrew J. Manson ◽  
Andrew Mounsey ◽  
Frederick J. Rowell ◽  
John Seviour
Keyword(s):  
Immunosorbent Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
End Point
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